摘要
采用反转录－聚合酶链反应（ＲＴ－ＰＣＲ），从１株传染性法氏囊病病毒（ＩＢＤＶ）本地分离株ＧＺ９０２中扩增到编码ＩＢＤＶ主要免疫蛋白ＶＰ２的ｃＤＮＡ片断，大小约为１３５０ｂｐ。将该ｃＤＮＡ片断插入ｐＢｓＫ质粒中，用重组质粒转化大肠杆菌ＸＬ１－Ｂｌｕｅ，成功获得重组质粒转化菌。对重组质粒ＤＮＡ进行酶切鉴定，证明插入的ＤＮＡ片断与ＰＣＲ扩增的ＤＮＡ片段大小相符。对ＧＺ９０２高可变区进行了序列分析。其核苷酸序列与３株ＩＢ－ＤＶ变异株Ａ、ＧＬＳ、Ｅ的同源性分别达到９８．９％，９６．２％和９５．５％。而与其他Ⅰ型毒株的同源性较低。比较从ＤＮＡ序列推导出的氨基酸序列，发现ＧＺ９０２高可变区的两个亲水区均发生了一个氨基酸的变化，而在２４９和２５４位上的氨基酸分别为Ｋ和Ｓ，与以上３个变异株相同，但与所有标准Ⅰ型毒株不同。这两个氨基酸可以作为ＩＢＤＶ变异株的标志。
Using Reverse Transcription Polymerase Chain Reaction (RT PCR) technique,the cDNA fragment with length of about 1350bp coding for immunodominant protein VP 2 of infectious bursal disease virus (IBDV) was amplified from Gz902,a variant strain isolated from Guangdong Province.The fragment digested with BgII and EcoRI were inserted into the BamHI/EcoRI sites of pBluescriptII KS vector,and the recombinant plasmid were transformed into E.coli strain XLI Blue.The positive recombinant colonies were identified by PCR and restriction endonuclease digestion.The nucleotide sequence of hypervariable region of VP 2 gene was analyzed using Autocycle DNA Sequencing kit.The result showed that the similarities of the DNA sequence of Gz902 strain with variant strain A,GLS and E are 98.9%,96.2%and 95.5% respectively.Comparing the deduced amino acid sequences,there is one amino acid changed in each of the two hydrophilc regions of Gz902 strain.At two positions (249 and 254),the amino acid residues of Gz902 strain are conserved in variant strain A,GLS and E,but differs from that of all the classical strains.These two amino acid residues can be regarded as the signature of variant strains of IBDV.
出处
《中国兽医杂志》
CAS
北大核心
1997年第9期3-5,共3页
Chinese Journal of Veterinary Medicine
关键词
变异株
基因克隆
序列分析
IBDV
Infectious bursal disease virus (IBDV) Variant strain Gene cloning DNA sequence
