摘要
以AcM NPV egt基因为探针,通过Southern 杂交和原位杂交,克隆了含EoSNPV egt基因的4.95 kb EcoRⅠ-K 片段,并进一步亚克隆了含完整egt基因编码区的1.8 kb HindⅢ-EcoRⅤ片段。应用双脱氧链终止法,测定了基因编码区中的XhoⅠ-HincⅡ片段471 bp 的核苷酸全序列。计算机分析表明,该片段编码的157 个氨基酸序列与其它杆状病毒egt基因保守区的Ⅳ(部分),Ⅴ,Ⅵ和Ⅶ区同源性很高,与BsSNPV,M bMNPV,LdM NPV,CfMNPV,AcMNPV,SlMNPV 基因相应序列的同源性分别为64.1% ,62.8% ,58.3% ,47.4% ,46.8% 和47.2% 。
EoSNPV 4.95 kb Eco RⅠ K fragment which contains the egt gene was cloned by Southern bloting and site hybridization methods, using the 32 P labled AcMNPV egt gene as a probe. The 1.8 kb Hind Ⅲ Eco RⅤ fragment containing the complete coding region of the egt gene was further subcloned. 471 nucleotide sequence ( Xho Ⅰ Hinc Ⅱ) in the coding region was sequenced with the dideoxynucleotide chain terminating method. Analysis with MALIGN software showed that 157 aa sequence encoded by this region of EoSNPV shares a high degree of homology with the conserved region IV(partial), V, VI and VII in EGTs of BsSNPV, MbMNPV, LdMNPV, CfMNPV, AcMNPV and SlMNPV, with 64.1%, 62.8%, 58.3%, 47.4%, 46.8%和47.2% amino acid identities, respectively.
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
1999年第6期583-588,共6页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
浙江省自然科学基金
关键词
茶尺蠖核型
多角体病毒
EGT基因
克隆
Ectropis obliqua single nucleocapsid nucleopolyhedrovirus
egt gene
cloning
nucleotide sequence
