银染色测定粘虫核多角体病毒多角体基因序列  

SEQUENCING OF POLYHEDRIN GENE OF LEUCANIA SEPARATA MULTIPLE CAPSID NUCLEAR POLYHEDROSIS VIRUS BY SILVER SEQUENCING SYSTEM

作　　者：王家旺[1] 黄永秀[1] 齐义鹏[1] 杨复华[1] 

机构地区：[1]武汉大学病毒学研究所

出　　处：《微生物学报》1995年第5期327-335,共9页Acta Microbiologica Sinica

基　　金：国家自然科学基金资助的课题

摘　　要：LsMNPV DNA用EcoRV酶切进行基因组克降,用AcMNPV的部分多角体基因顺序DNA片段作探针,菌落原位杂交法结合测序筛选到分别含LsMNPV部分多角体基因的重组质粒pLsEV1和pLsPH5。用银染色PCR线性扩增双脱氧法测序,发现LsMNPv的完整基因即位于这两个片段上。LsMNPV多角体基因长741bp,编码区碱基同源性与AcMNPV和MbMNPV分别为80.0％和97.0％,氨基酸同源性分别为89.8和97.5％。氨基酸组成中以谷氨酸(Glu)含量最高,谷氨酰胺和色氨酸含量最低。密码子选用以第三个碱基为嘧啶的密码子频率最高。多角体蛋白N端有一类似信号肽结构的26个氨基酸的疏水区。The genome of LsNPV was digested with EcoRV and cloned into pBlu-escript. With the partial polyhedrin gene of Autographa californica as a probe, positive recombinants were screened by colony in situ hybridization, further characterized by sequencing, and designated as pLsEV and pLsPH5. The intact 741 bp polyhedrin gene of LsNPV is located on the two recombinants revealed by Silver Sequencing System, and shares 80.0% and 97.0% nucleotide identity of the coding sequence, 87.8% and 97.5% amino acid identity with AcMNPV and MbMNPV respectively. Glu is the richest amino acid, and Gln and Trp the poorest amino acid in the polyhedrin of LsNPV. The codons ended with pyrimidine were higher in the codon usage. There was a signal-peptide-like hydrophobic region of 26 amino acids in the N end of polyhedrin.

关 键 词：粘虫 核型多角体病毒 昆虫病毒 多角体基因 

分 类 号：Q939.403[生物学—微生物学] S476.13[农业科学—农业昆虫与害虫防治]