家蚕核多角体病毒的多角体蛋白基因

本文用AcMNPV多角体蛋白质基因的克隆DNA作探针,从BmS NPV DNASal Ⅰ酶酶解片段基因库中筛选出一个有同源顺序的克隆pBN 61。该克隆含大小为1.65 kb插入DNA片段。本文报道了该片段的Hind Ⅲ,Hpa Ⅱ和Alu Ⅰ等限制性内切核酸酶酶切图谱分析和部分DNA顺序分析结果。测得的DNA顺序中有138个碱基与已报道的BmSNPV多角体蛋白质一级结构中的46个氨基酸顺序相比较,除一个氨基酸发生变化外,其余完全相一致。克隆pBN 61可能包含完整的Bm SNPV多角体蛋白质结构基因。     

THE CLONED SALI FRAGMENTS OF Bombyx mori NUCLEAR POLYHEDROSIS VIRUS DNA

Bombyx mori nuclear polyhedrosis virus (BmNPV) DNA has been cleaved with restriction endonuclease SalI into 29 fragments of different sizes. The range of the molecular weight of these fragments as detected by electrophoresis on agarose gel is from 0.70 Kb to 10.0Kb. 24 SalI fragments were cloned in plasmid pBR322. The sum of the molecular weight of cloned BmNPV DNA fragments accounts for about 80% of the virus genome DNA.     

THE POLYHEDRIN GENE OF Bombyx mori NUCLEAR POLYHEDROSIS VIRUS

Cloned SalI fragments of Bombyx mori nuclear polyhedrosis virus (BmSNPV) DNA were screened with the polyhedrin gene of Autographa californica nuclear polyhedrosis virus as a probe. One positive clone, pBN61, with an insert of 1.65 Kb, was obtained. The Hind-Ⅲ, HpaⅡ and AluⅠ maps of the insert were constructed. Part of its nucleotide sequence has been determined. The 46 amino acid sequence, as determined from the nucleotide sequence, was compared with the reported sequence of BmSNPV polyhedrin. Only ono amino acid difference has been found. It is likely that clone pBN61 contains the whole BmSNPV polyhedrin gene.     

家蚕核多角体病毒基因库

本文报道家蚕核多角体病毒基因组DNA经限制性内切酶SalI酶解,琼脂糖凝胶电泳分离,得0.70至 10.0kb大小不同的29种片段.所得 DNA片段与 SalI酶解之质粒pBR 322 DNA进行体外重组后,经转化大肠杆菌 HB 101菌株,获得带重组质粒克隆株.根据重组质粒DNA 中SalI酶插八片段的分子量、Southern法DNA杂交及多种限制性内切酶酶切点等方法鉴定,证明已将家蚕核多角体病毒DNA的24种不同大小的片段克隆在质粒 pBR 322 中.克隆的 DNA片段总长度占病毒基因组DNA的百分之八十。     

DNA FRAGMENTS OF Bombyx mori NUCLEAR POLYHEDROSIS VIRUS CONTAINING THE PROMOTERS ACTIVE IN PROKARYOTES

Using a promoter probe plasmid in E. coli called pHE5, eight different HindⅢ and one SalⅠDNAfragments of Bombyx mori nuclear polyhedrosis virus, directing the expression of the tetracycline re-sistance gene, have been cloned and isolated. The tetracycline resistance levels of the strains containing therecombinant plasmids were measured. Among them, the highest level of the resistance to tetracyclinewas 30μg/ml. Part of the nucleotide sequence of a DNA fragment was determined. A sequence similarto the E. coli promoter was found.