[Genetic analysis of NS1 fragment of human H5N1 influenza virus isolated in Anhui province and its expression in Escherichia coli].
NS1 gene was amplified from an H5N1 influenza virus, A/Anhui/01/2005 for cloning, sequence analysis and later expression in Escherichia coli. The result showed that NS1 of A/Anhui/01/2005 strain had the close phylogenetic relationship with that of H5N1 avian influenza strains recently isolated in Fujian and Hunan. Correspondingly, its amino acid sequence showed the highest homology with that of Fujian and Hunan strains. The amino acid position of 92 involved in virus virulence was Asp, contrast to Glu in A/HK/156/97. Five amino acid deletion from 80 to 84 was also found in A/Anhui/01/2005, which was considered as a contributor to virus resistance against cytokine, such as IFN, TNF, etc. A motif binding to CBSF, conveted to GFWEN, which was different from previous GLEWN found in other 19 strains. Besides, the PL motif of ESEV, binding to PDZ domain, is the same as previous high-mortality H5N1 isolates. Furthermore, this NS1 was efficiently expressed in Escherichia coli and the highly purified product demonstrated wonderful activity as confirmed by Western blot. As a result, the work paves the way for further understanding the role of NSI in human H5N1 infection and development of new antiviral drugs against influenza virus.
Cheng
Cong-sheng
CS
National Key Laboratory for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China. cshengc@hotmail.com
Lan
Yu
Y
Liu
Yan
Y
Zhang
Zhi-qing
ZQ
Shu
Yue-long
YL
D002681:China; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D007251:Influenza, Human / Q000821:virology*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
C085820:INS1 protein, influenza virus; D017361:Viral Nonstructural Proteins