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人工合成引物Y_5和Y_3,在引物Y_5中引入Nco I位点,通过PCR技术扩增了中国株系PVY Nib基因.DNA全序列测定的结果表明,该基因由1557个核苷酸组成,编码521个氨基酸.与PVY的N株系和GO16株系相应基因之间的核苷酸同源性为91.12％和80.06％,氨基酸的同源性为94.44％和91.3％.对已报道的马铃薯Y病毒组中的9个病毒Nib基因的氨基酸序列进行了比较分析,发现它们均有8个保守区以及GDD和NTP结构.将全长Nib基因重组到表达载体pTricHis(B)中并在大肠杆菌中得到了表达.
The primers Y_5 and Y_3 were synthesized and the Nco I site was introduced into the primer Y_5 to amplify the PVY Nib gene from China by PCR.The results of DNA sequencing showed that the gene consisted of 1557 nucleotides encoding 521 Amino acids.The nucleotide homologies with the corresponding genes of PVY N strain and GO16 strain were 91.12% and 80.06%, and the amino acid homologies were 94.44% and 91.3% The amino acid sequences of 9 virus Nib genes in the group were compared and found that they all have 8 conserved regions and GDD and NTP structures.The full length Nib gene was recombined into the expression vector pTricHis (B) and obtained in E. coli The expression.