Research Article | Volume: 5, Issue: 5, Sep-Oct, 2017

Rational Design of Duplex Specific Nuclease for One-Step Isothermal Viral RNA Detection

Elizabeth M. Wurtzler Ranjani Ravi Vikram Kapoor David Wendell   

Open Access   

Published:  Oct 30, 2017

DOI: 10.7324/JABB.2017.50501
Abstract

RNA viruses are a potent human adversary, evidenced by several global pandemics including the Ebolavirus in West Africa, the emerging Zika virus, and outbreaks of new Influenza strains and Norwalk virus in the food supply and cruise ships. Despite the virulence of these pathogens, there remains a significant limitation for detecting these viruses in a fast, accurate and cost effective manner. To meet this need we present a modified form of the duplex specific nuclease enzyme from the Paralithodes camtschaticus crab capable of generating an RNA-based signal amplification in a fraction of the time required for standard RT-qPCR. The applicability of this enzyme is demonstrated in an assay for Norwalk virus detection with a lower limit of ~100 viral copies per liter of environmental water.


Keyword:     Duplex specific nuclease norovirus viral detection RNA RNA detection environmental water.


Citation:

Wurtzler EM, Ravi R, Kapoor V, Wendell D. Rational Design of Duplex Specific Nuclease for One-Step Isothermal Viral RNA Detection. J App Biol Biotech. 2017; 5 (05): 1-10.

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

HTML Full Text
Reference

1. D. R. Walt. Miniature Analytical Methods for Medical Diagnostics. Science. 2005; 308(5719): 217-219.

2. D. A. Relman. The search for unrecognized pathogens. Science. 1999; 284(5418): 1308-1310.

3. J. Lamb, E. D. Crawford, D. Peck, J. W. Modell, I. C. Blat, M. J. Wrobel, J. Lerner, J.-P. Brunet, A. Subramanian, K. N. Ross, M. Reich, H. Hieronymus, G. Wei, S. A. Armstrong, S. J. Haggarty, P. A. Clemons, R. Wei, S. A. Carr, E. S. Lander,T. R. Golub. The Connectivity Map: Using Gene-Expression Signatures to Connect Small Molecules, Genes, and Disease. Science. 2006; 313(5795): 1929-1935.

4. J. Drews. Drug Discovery: A Historical Perspective. Science. 2000; 287(5460): 1960-1964.

5. E. DeLong, G. Wickham,N. Pace. Phylogenetic stains: ribosomal RNA-based probes for the identification of single cells. Science. 1989; 243(4896): 1360-1363.

6. M. Mandal,R. R. Breaker. Gene regulation by riboswitches. Nat Rev Mol Cell Biol. 2004; 5(6): 451-463.

7. J. Compton. Nucleic acid sequence-based amplification. Nature. 1991; 350(6313): 91-92.

8. Mader, U. Riehle, T. Brandstetter, E. Stickeler, A. zur Hausen,J. Rühe. Microarray-based amplification and detection of RNA by nucleic acid sequence based amplification. Analytical and Bioanalytical Chemistry. 2010; 397(8): 3533-3541.

9. Heim, I. M. Grumbach, S. Zeuke,B. Top. Highly sensitive detection of gene expression of an intronless gene: amplification of mRNA, but not genomic DNA by nucleic acid sequence based amplification (NASBA). Nucleic Acids Research. 1998; 26(9): 2250-2251.

10. Lagunavicius, E. Merkiene, Z. Kiveryte, A. Savaneviciute, V. Zimbaite-Ruskuliene, T. Radzvilavicius,A. Janulaitis. Novel application of Phi29 DNA polymerase: RNA detection and analysis in vitro and in situ by target RNA-primed RCA. RNA. 2009; 15(5): 765-771.

11. E. Merkiene, E. Gaidamaviciute, L. Riauba, A. Janulaitis,A. Lagunavicius. Direct detection of RNA in vitro and in situ by target-primed RCA: The impact of E. coli RNase III on the detection efficiency of RNA sequences distanced far from the 3′-end. RNA. 2010; 16(8): 1508-1515.

12. T. Notomi, H. Okayama, H. Masubuchi, T. Yonekawa, K. Watanabe, N. Amino,T. Hase. Loop-mediated isothermal amplification of DNA. Nucleic Acids Research. 2000; 28(12): e63-e63.

13. T. C. Cardoso, H. F. Ferrari, L. C. Bregano, C. Silva-Frade, A. C. G. Rosa,A. L. Andrade. Visual detection of turkey coronavirus RNA in tissues and feces by reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with hydroxynaphthol blue dye. Molecular and Cellular Probes. 2010; 24(6): 415-417.

14. Y. Zhao, L. Zhou,Z. Tang. Cleavage-based signal amplification of RNA. Nat Commun. 2013; 4(1493.

15. V. E. Anisimova, D. V. Rebrikov, D. A. Shagin, V. B. Kozhemyako, N. I. Menzorova, D. B. Staroverov, R. Ziganshin, L. L. Vagner, V. A. Rasskazov, S. A. Lukyanov,A. S. Shcheglov. Isolation, characterization and molecular cloning of duplex-specific nuclease from the hepatopancreas of the Kamchatka crab. BMC Biochem. 2008; 9(14.

16. D. A. Shagin, D. V. Rebrikov, V. B. Kozhemyako, I. M. Altshuler, A. S. Shcheglov, P. A. Zhulidov, E. A. Bogdanova, D. B. Staroverov, V. A. Rasskazov,S. Lukyanov. A novel method for SNP detection using a new duplex-specific nuclease from crab hepatopancreas. Genome Res. 2002; 12(12): 1935-1942.

17. B.-C. Yin, Y.-Q. Liu,B.-C. Ye. One-Step, Multiplexed Fluorescence Detection of microRNAs Based on Duplex-Specific Nuclease Signal Amplification. Journal of the American Chemical Society. 2012; 134(11): 5064-5067.

18. P. A. Zhulidov, E. A. Bogdanova, A. S. Shcheglov, L. L. Vagner, G. L. Khaspekov, V. B. Kozhemyako, M. V. Matz, E. Meleshkevitch, L. L. Moroz, S. A. Lukyanov,D. A. Shagin. Simple cDNA normalization using kamchatka crab duplex-specific nuclease. Nucleic Acids Research. 2004; 32(3): e37-e37.

19. E. A. Bogdanova, E. V. Barsova, I. A. Shagina, A. Scheglov, V. Anisimova, L. L. Vagner, S. A. Lukyanov,D. A. Shagin. Normalization of full-length-enriched cDNA. Methods Mol Biol. 2011; 729(85-98.

20. D. C. Christodoulou, J. M. Gorham, D. S. Herman,J. G. Seidman. Construction of normalized RNA-seq libraries for next-generation sequencing using the crab duplex-specific nuclease. Curr Protoc Mol Biol. 2011; Chapter 4(Unit4 12.

21. Shagina, E. Bogdanova, I. Z. Mamedov, Y. Lebedev, S. Lukyanov,D. Shagin. Normalization of genomic DNA using duplex-specific nuclease. Biotechniques. 2010; 48(6): 455-459.

22. E. A. Bogdanova, I. A. Shagina, E. Mudrik, I. Ivanov, P. Amon, L. L. Vagner, S. A. Lukyanov,D. A. Shagin. DSN depletion is a simple method to remove selected transcripts from cDNA populations. Molecular Biotechnology. 2009; 41(3): 247-253.

23. E. A. Bogdanova, I. A. Shagina, Y. G. Yanushevich, L. L. Vagner, S. A. Lukyanov,D. A. Shagin. Preparation of prokaryotic cDNA for full-scale transcriptome analysis. Russian Journal of Bioorganic Chemistry. 2011; 37(6): 775-778.

24. H. Yi, Y. J. Cho, S. Won, J. E. Lee, H. Jin Yu, S. Kim, G. P. Schroth, S. Luo,J. Chun. Duplex-specific nuclease efficiently removes rRNA for prokaryotic RNA-seq. Nucleic Acids Res. 2011; 39(20): e140.

25. R. H. Peng, A. S. Xiong, Y. Xue, X. Li, J. G. Liu, B. Cai,Q. H. Yao. Kamchatka crab duplex-specific nuclease-mediated transcriptome subtraction method for identifying long cDNAs of differentially expressed genes. Anal Biochem. 2008; 372(2): 148-155.

26. F. Swennenhuis, B. Foulk, F. A. W. Coumans,L. W. M. M. Terstappen. Construction of repeat-free fluorescence in situ hybridization probes. Nucleic Acids Research. 2012; 40(3): e20-e20.

27. Y. Zhao, H. Hoshiyama, J. W. Shay,W. E. Wright. Quantitative telomeric overhang determination using a double-strand specific nuclease. Nucleic Acids Research. 2008; 36(3): e14-e14.

28. Y. Zhao, J. W. Shay,W. E. Wright. Telomere G-overhang length measurement method 1: the DSN method. Methods Mol Biol. 2011; 735(47-54.

29. M. Liu, M. Yuan, X. Lou, H. Mao, D. Zheng, R. Zou, N. Zou, X. Tang,J. Zhao. Label-free optical detection of single-base mismatches by the combination of nuclease and gold nanoparticles. Biosens Bioelectron. 2011; 26(11): 4294-4300.

30. Z. Kapikian, R. G. Wyatt, R. Dolin, T. S. Thornhill, A. R. Kalica,R. M. Chanock. Visualization by Immune Electron Microscopy of a 27-nm Particle Associated with Acute Infectious Nonbacterial Gastroenteritis. Journal of Virology. 1972; 10(5): 1075-1081.

31. P. R. Lambden, E. O. Caul, C. R. Ashley,I. N. Clarke. Sequence and genome organization of a human small round-structured (Norwalk-like) virus. Science. 1993; 259(5094): 516-519.

32. P. R. Lambden, B. Liu,I. N. Clarke. A conserved sequence motif at the 5' terminus of the Southampton virus genome is characteristic of the Caliciviridae. Virus Genes. 1995; 10(2): 149-152.

33. X. Jiang, M. Wang, K. Wang,M. K. Estes. Sequence and genomic organization of Norwalk virus. Virology. 1993; 195(1): 51-61.

34. N. Xi, D. Y. Graham, K. N. Wang,M. K. Estes. Norwalk virus genome cloning and characterization. Science. 1990; 250(4987): 1580-1583.

35. R. L. Fankhauser, S. S. Monroe, J. S. Noel, C. D. Humphrey, J. S. Bresee, U. D. Parashar, T. Ando,R. I. Glass. Epidemiologic and molecular trends of "Norwalk-like viruses" associated with outbreaks of gastroenteritis in the United States. J Infect Dis. 2002; 186(1): 1-7.

36. S. M. Green, P. R. Lambden, E. O. Caul, C. R. Ashley,I. N. Clarke. Capsid diversity in small round-structured viruses: molecular characterization of an antigenically distinct human enteric calicivirus. Virus Res. 1995; 37(3): 271-283.

37. S. M. Karst, C. E. Wobus, M. Lay, J. Davidson,H. W. t. Virgin. STAT1-dependent innate immunity to a Norwalk-like virus. Science. 2003; 299(5612): 1575-1578.

38. S. L. Oliver, A. M. Dastjerdi, S. Wong, L. El-Attar, C. Gallimore, D. W. Brown, J. Green,J. C. Bridger. Molecular characterization of bovine enteric caliciviruses: a distinct third genogroup of noroviruses (Norwalk-like viruses) unlikely to be of risk to humans. J Virol. 2003; 77(4): 2789-2798.

39. J. Vinje,M. P. Koopmans. Simultaneous detection and genotyping of "Norwalk-like viruses" by oligonucleotide array in a reverse line blot hybridization format. J Clin Microbiol. 2000; 38(7): 2595-2601.

40. de Wit, M. P. Koopmans, L. M. Kortbeek, W. J. Wannet, J. Vinje, F. van Leusden, A. I. Bartelds,Y. T. van Duynhoven. Sensor, a population-based cohort study on gastroenteritis in the Netherlands: incidence and etiology. Am J Epidemiol. 2001; 154(7): 666-674.

41. E. Scallan, R. M. Hoekstra, F. J. Angulo, R. V. Tauxe, M. A. Widdowson, S. L. Roy, J. L. Jones,P. M. Griffin. Foodborne illness acquired in the United States--major pathogens. Emerg Infect Dis. 2011; 17(1): 7-15.

42. M. Patel, M. A. Widdowson, R. I. Glass, K. Akazawa, J. Vinje,U. D. Parashar. Systematic literature review of role of noroviruses in sporadic gastroenteritis. Emerg Infect Dis. 2008; 14(8): 1224-1231.

43. O’Ryan, M. Riera-Montes, B. Lopman. Norovirus in Latin America: Systematic Review and Meta-Analysis. Pediatr Infect Dis J. 2016; Published Ahead of Print.

44. K. Pringle, B. Lopman, E. Vega, J. Vinje, U. Parashar, A. Hall. Noroviruses: epidemiology, immunity and prospects for prevention. Future Microbiology. 2015; 10(1): 53-67.

45. F. Kowalzik, M. Riera-Montes, T. Verstraeten, F. Zepp. The Burden of Norovirus Disease in Children in the European Union. 2015. 34(3): 229-234

46. J. Mans, G. E. Armah, A. D. Steele, M. B. Taylor. Norovirus Epidemiology in Africa: A Review. PLoS ONE, 2016; 11(4): e0146280.

47. S. M. Ahmed, A. J. Hall, A. E. Robinson, L. Verhoef, P. Premkumar, U. D. Parashar, M. Koopmans,B. A. Lopman. Global prevalence of norovirus in cases of gastroenteritis: a systematic review and meta-analysis. The Lancet Infectious Diseases. 2014; 14(8): 725-730.

48. Lopman, A. J. Hall, A. T. Curns,U. D. Parashar. Increasing rates of gastroenteritis hospital discharges in US adults and the contribution of norovirus, 1996-2007. Clin Infect Dis. 2011; 52(4): 466-474.

49. J. Hall, A. T. Curns, L. C. McDonald, U. D. Parashar,B. A. Lopman. The roles of Clostridium difficile and norovirus among gastroenteritis-associated deaths in the United States, 1999-2007. Clin Infect Dis. 2012; 55(2): 216-223.

50. F. Teunis, C. L. Moe, P. Liu, S. E. Miller, L. Lindesmith, R. S. Baric, J. Le Pendu,R. L. Calderon. Norwalk virus: how infectious is it? J Med Virol. 2008; 80(8): 1468-1476.

51. T. Halperin, M. Yavzori, A. Amitai, E. Klement, R. Kayouf, I. Grotto, M. Huerta, L. A. Hadley, S. S. Monroe, D. Cohen,N. Orr. Molecular analysis of noroviruses involved in acute gastroenteritis outbreaks in military units in Israel, 1999-2004. Eur J Clin Microbiol Infect Dis. 2005; 24(10): 697-700.

52. E. L. Yee, H. Palacio, R. L. Atmar, U. Shah, C. Kilborn, M. Faul, T. E. Gavagan, R. D. Feigin, J. Versalovic, F. H. Neill, A. L. Panlilio, M. Miller, J. Spahr,R. I. Glass. Widespread outbreak of norovirus gastroenteritis among evacuees of Hurricane Katrina residing in a large "megashelter" in Houston, Texas: lessons learned for prevention. Clin Infect Dis. 2007; 44(8): 1032-1039.

53. L. Verhoef, E. Depoortere, I. Boxman, E. Duizer, Y. van Duynhoven, J. Harris, C. Johnsen, A. Kroneman, S. Le Guyader, W. Lim, L. Maunula, H. Meldal, R. Ratcliff, G. Reuter, E. Schreier, J. Siebenga, K. Vainio, C. Varela, H. Vennema,M. Koopmans. Emergence of New Norovirus Variants on Spring Cruise Ships and Prediction of Winter Epidemics. Emerging Infectious Disease journal. 2008; 14(2): 238.

54. S. R. Seitz, J. S. Leon, K. J. Schwab, G. M. Lyon, M. Dowd, M. McDaniels, G. Abdulhafid, M. L. Fernandez, L. C. Lindesmith, R. S. Baric,C. L. Moe. Norovirus Infectivity in Humans and Persistence in Water. Applied and Environmental Microbiology. 2011; 77(19): 6884-6888.

55. J. L. Hyde, S. V. Sosnovtsev, K. Y. Green, C. Wobus, H. W. Virgin,J. M. Mackenzie. Mouse norovirus replication is associated with virus-induced vesicle clusters originating from membranes derived from the secretory pathway. J Virol. 2009; 83(19): 9709-9719.

56. S. Taube, A. Kolawole, M. Höhne, J. Wilkinson, S. Handley, J. Perry, L. Thackray, R. Akkina, C.Wobus.A Mouse Model for Human Norovirus.mBio. 2013; 4(4): e00450-13

57. Khamrin, A. Thongprachum, S. Okitsu, N. Maneekarn, S. Hayakawa,H. Ushijima. Comparison of three rapid tests for detection of norovirus in stool samples of acute gastroenteritis pediatric patients. Journal of Tropical Pediatrics. 2014; 60(6): 481-483.

58. Derrington, F. Moore, D. Thorley,T. Constantino. Evaluation of Immunochromatographic Test Kits For the Detection of Norovirus in Faecal Specimens. Pathology - Journal of the RCPA. 2014; 46(S100).

59. L. Atmar,M. K. Estes. Diagnosis of noncultivatable gastroenteritis viruses, the human caliciviruses. Clin Microbiol Rev. 2001; 14(1): 15-37.

60. M. S. Plantenga, B. Shiferaw, W. E. Keene, C. Biggs, J. M. Terry, L. Grenz,P. R. Cieslak. Specimen Collection and Confirmation of Norovirus Outbreaks. Emerging Infectious Diseases. 2011; 17(8): 1553-1555.

61. H. C. P. Santos, L. C. Turones, F. S. Fiaccadori, D. d. D. de Paula Cardoso,M. B. de Lima Dias. Screening of Fecal Samples From Asymptomatic Children, For Norovirus Detection, Using a Third Generation Enzyme Immunoassay Commercial Kit. Revista de Patologia Tropical. 2014; 43(2): 143-149.

62. G. Lennon, N. Reidy, P. J. Collins, L. Gunn, P. V. Coyle, B. Cryan, S. Fanning,H. O’Shea. A comparison of the efficiency of ELISA and selected primer sets to detect Norovirus isolates in southern Ireland over a four-year period (2002-2006): variation in detection rates and evidence for continuing predominance of NoV GII.4 genotype. Archives of Virology. 2014; 159(7): 1697-1705.

63. F.-R. Lin, Y.-H. Shen, C.-W. Fang, S.-S. Shie, C.-G. Huang, S. Yang, S.-L. Yang, K.-C. Tsao, Y.-C. Huang, M.-W. Lai,C.-J. Chen. Incidence of and Factors Associated with False Positives in Laboratory Diagnosis of Norovirus Infection by Amplification of the RNA-Dependent RNA Polymerase Gene. PLoS ONE. 2014; 9(9): e109876.

64. N. L. Dunbar, L. D. Bruggink,J. A. Marshall. Evaluation of the RIDAGENE real-time PCR assay for the detection of GI and GII norovirus. Diagn Microbiol Infect Dis. 2014; 79(3): 317-321.

65. K. Ambert-Balay, P. Pothier. Evaluation of 4 immuno-chromatographic tests for rapid detection of norovirus in faecal samples. Journal of Clinical Virology. 2013; 56(3): 278-282.

66. V. Costantini, L. Grenz, A. Fritzinger, D. Lewis, C. Biggs, A. Hale,J. Vinjé. Diagnostic Accuracy and Analytical Sensitivity of IDEIA Norovirus Assay for Routine Screening of Human Norovirus. Journal of Clinical Microbiology. 2010; 48(8): 2770-2778.

67. N. Shigemoto, Y. Tanizawa, T. Matsuo, N. Sakamaki, Y. Ohiro, S. Takayasu,S. Fukuda. Clinical evaluation of a bioluminescent enzyme immunoassay for detecting norovirus in fecal specimens from patients with acute gastroenteritis. Journal of Medical Virology. 2014; 86(7): 1219-1225.

68. K. Hanaki, F. Ike, A. Kajita, W. Yasuno, M. Yanagiba, M. Goto, K. Sakai, Y. Ami,S. Kyuwa. Detection of murine norovirus by reverse transcription loop-mediated isothermal amplification. J Virol Methods. 2014; 204(17-24.

69. J. Luo, Z. Xu, K. Nie, X. Ding, L. Guan, J. Wang, Y. Xian, X. Wu,X. Ma. Visual detection of norovirus genogroup II by reverse transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye. Food Environ Virol. 2014; 6(3): 196-201.

70. Hong, J. Kwon, D. Kim,S. Yang. A rapid, sensitive and selective electrochemical biosensor with concanavalin A for the preemptive detection of norovirus. Biosensors and Bioelectronics. 2015; 64(0): 338-344.

71. J. Pratt, I.J. MacRae. The RNA-induced Silencing Complex: A Versatile Gene-silencing Machine. Journal of Biological Chemistry. 2009; 284(27): 17897-17901.

72. L. Hofacker, W. Fontana, P. F. Stadler, L. S. Bonhoeffer, M. Tacker,P. Schuster. Fast folding and comparison of RNA secondary structures. Monatshefte für Chemie / Chemical Monthly. 1994; 125(2): 167-188.

73. M. Hoehne,E. Schreier. Detection of Norovirus genogroup I and II by multiplex real-time RT- PCR using a 3'-minor groove binder-DNA probe. BMC Infectious Diseases. 2006; 6(69-69.

74. M. L. Stewart, A. P. Grollman,M.-T. Huang. Aurintricarboxylic acid: inhibitor of initiation of protein synthesis. Proceedings of the National Academy of Sciences. 1971; 68(1): 97-101.

75. Y. Yap, X. Zhang, A. Andonov,R. He. Structural analysis of inhibition mechanisms of Aurintricarboxylic Acid on SARS-CoV polymerase and other proteins. Computational Biology and Chemistry. 2005; 29(3): 212-219.

76. G. J. Bartlett, C. T. Porter, N. Borkakoti, J. M. Thornton. Analysis of catalytic residues in enzyme active sites. Journal of Molecular Biology. 2002; 324(1): 105-121.

77. Vogelstein, K. W. Kinzler. Digital PCR. Proceedings of the National Academy of Sciences. 1999; 96(16): 9236-9241.

78. Bustin. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol. 2000; 25(2): 169-193.

79. S. G. Lee, S. H. Lee, S. W. Park, C. I. Suh, W. H. Jheong, S. Oh,S. Y. Paik. Standardized positive controls for detection of norovirus by reverse transcription PCR. Virol J. 2011; 8(260.

80. R. I. Glass, U. D. Parashar, M. K. Estes. Norovirus Gastroenteritis. New England Journal of Medicine. 2009; 361(18): 1776-1785.

81. R. Brennan, B. W. Matthews. The helix-turn-helix DNA binding motif. J Biol Chem. 1989; 264(4): 1903-1906.

82. H. Ohlendorf, B. W. Matthews. Structural studies of protein-nucleic acid interactions. Annu Rev Biophys Bioeng. 1983; 12(259-284.

83. Y. Takeda, D. H. Ohlendorf, W. F. Anderson,B. W. Matthews. DNA-binding proteins. Science. 1983; 221(4615): 1020-1026.

84. O. Pabo, R. T. Sauer. Protein-DNA recognition. Annu Rev Biochem. 1984; 53(293-321.

85. R. Schleif. DNA binding by proteins. Science. 1988; 241(4870): 1182-1187.

86. R. Zhang, A. Joachimiak, C. Lawson, R. Schevitz, Z. Otwinowski, P. Sigler. The crystal structure of trp aporepressor at 1.8 A shows how binding tryptophan enhances DNA aflinity. Nature. 1987; 327(591-597.

87. Z. Otwinowski, R. Schevitz, R. Zhang, C. Lawson, A. Joachimiak, R. Marmorstein, B. Luisi, P. Sigler. Crystal structure of trp represser/operator complex at atomic resolution. Nature. 1988; 335(6188): 321-329.

88. Yang, R. Yan, A. Roy, D. Xu, J. Poisson,Y. Zhang. The I-TASSER Suite: protein structure and function prediction. Nature methods. 2015; 12(1): 7-8.

89. Roy, A. Kucukural, Y. Zhang. I-TASSER: a unified platform for automated protein structure and function prediction. Nature protocols. 2010; 5(4): 725-738.

90. Y. Zhang. I-TASSER server for protein 3D structure prediction. BMC bioinformatics. 2008; 9(1):40

Article Metrics

133 Absract views 175 PDF Downloads 308 Total views

Related Search

By author names

Citiaion Alert By Google Scholar


Similar Articles

Morphological, genetic and symbiotic characterization of root nodule bacteria isolated from Bambara groundnuts (Vigna subterranea L. Verdc) from soils of Lake Victoria basin, western Kenya

Onyango Benson , Anyango Beatrice, Nyunja Regina, P. K. Koech, Robert A. Skilton , Stomeo Francesca

cDNA cloning and analysis of alanine-rich cuticle protein gene of common fouling barnacle, Balanus amphitrite

Chandrakant B. Jagtap,Om Kinker Tiwari,Pradeep Kumar

Isolation and optimization of alkaline protease producing Bacteria from undisturbed soil of NE-region of India falling under Indo-Burma biodiversity hotspots

Onkar Nath Tiwari, Thiyam Bidyababy Devi, Kangjam Sarabati Devi, Gunapati Oinam, Thingujam Indrama, Keithellakpam Ojit, Oinam Avijeet, Lakreiphy Ningshen

Production of exopolysaccharides by the cyanobacterium Anabaena sp. BTA992 and application as bioflocculants

Romi Khangembam, Onkar Nath Tiwari, Mohan Chandra Kalita

Molecular Phylogeny of Balsams (Genus Impatiens) Based on ITS Regions of Nuclear Ribosomal DNA Implies Two Colonization Events in South India

P. P. Shajitha, N. R. Dhanesh, P. J. Ebin, Laly Joseph, Aneesha Devassy, Reshma John, Jomy Augustine, Linu Mathew

Identification and profiling of high temperature responsive miRNAs in French bean (Phaseolus vulgaris L)

M. N. Jyothi,, S. Usha,, B. Suchithra,, N. Sharadamma , D. V. Rai, V. R. Devaraj, R. Nagesh Babu

Recent developments in understanding the mechanism and functions of microRNAs

Suhail Muzaffar, Karishma Bisht

Computational identification of miRNAs and their targets from Niger (Guizotia abyssinica)

K. Y. Prathiba, S. Usha , B. Suchithra , M. N. Jyothi , V. R. Devaraj, R. Nageshbabu

Biochemical and molecular characterization of a new pullulan producer Rhodosporidium paludigenum PUPY-06

RS Singh, Navpreet Kaur

Bio-prospecting of marine-derived fungi with special reference to production of keratinase enzyme - A need-based optimization study

P. Samuel, M. Maheswari, J. Vijayakumar, T. Selvarathinam, K. Amirtharaj, R. Deenathayalan

Long Non-Coding RNAs and their “Orchestration” in Cancers

Venkata Narasimha Kadali, Sheji Chandran, Smitha Murthy

Progress in understanding the regulation and expression of genes during plant somatic embryogenesis: A review

Vikrant, Prajisha Janardhanan

Inulinase production in shake-flask fermentations from Mucor circinelloides BGPUP-9

Ram Sarup Singh, Kanika Chauhan, Randeep Kaur, Ramanpreet Kaur

Molecular detection and antimicrobial resistance of Clostridium perfringens isolated from diabetic patients and bullet wounds

Aliaa Mohamed Hmood, Maysa S M Al-Shukri, Alaa H Al-Charrakh

Genome complexity of begomovirus disease and a concern in agro-economic loss

Hanjabam Joykishan Sharma, Susheel Kumar Sharma, Nongthombam Bidyananda Singh

Identification and characterization of causative agents of brown leaf spot disease of cassava in Kenya

Perpetuar Wangari Ng’ang’a, Douglas Watuku Miano, John Maina Wagacha, Paul Kuria

In silico analysis of chili encoded miRNAs targeting Chili leaf curl begomovirus and its associated satellite

Megha Mishra, Rajesh Kumar, Rajarshi Kumar Gaur, Rakesh Kumar Verma

Efficacy of antibiotic sensitivity and antimicrobial activity of Streptomyces cinereoruber RSA-14 isolated from rhizosphere soil of Alternanthera sessilis (L.) R. Br. ex DC

Sreenivasa Nayaka, Halaswamy Hiremath, Bidhayak Chakraborty, Pallavi Sathyanarayan Swamy, Dhanyakumara Shivapoojar Basavarajappa, Shashiraj Kareyallappa Nagaraja, Muthuraj Rudrappa, Meghashyama Prabhakara Bhat, Dattatraya Airodagi, Meenakshi Shivanandagouda Murigennavar

Plant induced resistance in Solanacearum lycopersicum species against root knot nematodes

B. Navyashree, Chandan Dharmashekar, Chandan Shivamallu, S. Balasubramanian, Shashanka K. Prasad, Shiva Prasad K, K. C. Latha

An effective and eco-friendly technique for control of post-harvest fungal pathogens of orange (Citrus sinensis) isolated from the distribution chain of Delhi NCR

Gayatri Krishna, Geethu Gopinath, Anupama Sharma Avasthi

In vitro biocontrol scenarios of Bacillus amyloliquefaciens subsp. amyloliquefaciens strain RLS19 in response to Alternaria macrospora, an Alternaria leaf spot phytopathogen of Bt cotton

Laxman Shrirangrao Raut, Ravindra Raosaheb Rakh, Venkat Shankarrao Hamde

Floral induction pathways: Decision making and determination in plants to flower-A comprehensive review

Latif Ahmad Peer, Mohd Yaqub Bhat, Nusrat Ahmad, Bilal Ahmad Mir

Microstructure characterization of male and female external genitalia of soapberry bug, Leptocoris augur (Hemiptera: Rhopalidae)

Vaishali J. Badwaik, Deepak D. Barsagade

Identification of noval non-coding RNAs in Rhizoctonia solani through mining of transcriptomic data

Balachander Durairaj, Sudheesh K. Prabhudas, Jaiganesh Rengarajan, Iyappan Sellamuthu

Molecular identification of endophytic fungi associated with Coleus forskohlii (Willd.) Briq.

Grace Leena Crasta,, Koteshwar Anandrao Raveesha,

Augmentative role of Piriformospora indica fungus and plant growth promoting bacteria in mitigating salinity stress in Trigonella foenum-graecum

Sanskriti Bisht, Shatrupa Singh, Madhulika Singh,, Jai Gopal Sharma

Expression of FUB-1 and FUB-11 as Toxic genes responsible for virulence during pathogenesis and combination of biocontrol agents in inhibition of Fusaric acid of Fusarium oxysporum causing Fusarium wilt of Arachis hypogaea L.

Pilli Rajeswari

Diversity due to mutations in circulating virus strains of SARS-CoV-2 may delay control of COVID-19

Sharanagouda S. Patil, Chandan Shivamallu, Chandan Dharmashekara, Sushma Pradeep, Kuralayanapalya Puttahonnappa Suresh, Ashwini Prasad, Shiva Prasad Kollur, Mahendra P. Yadav, Chandrashekar Srinivasa, Bramhadev Pattnaik

Role of microRNAs in the progression and metastasis of gastric cancer

Havisha Dinesh, Megala Jayaraman

Identification, production, and purification of a novel lipase from Bacillus safensis

Krishna Patel, Samir Parikh

Isolation and characterization of starch degrading bacteria from disparate soil samples

Vidhyutha Srivathsan, Mahima Bhandari, Priya Swaminathan

Exosome mediated cell signal toward breast cancer metastasis: A comprehensive review

Manikantan Pappuswamy, Anushka Shitut

Designing of guide RNA constructs for CRISPR/Cas9-mediated editing of rice transcription factor OsMADS26 for enhancing drought tolerance

K. Anjala, Rehna Augustine