摘要
本文报道了柞蚕核型多角体病毒(Anlherdea peruyi Nuclear polybedrosis Virus,ApNPV)DNA的分离、纯化,ApNPV DNA用限制性内切酶PstI、BamHI酶解分别出现31、6个片段。用Southern转印杂交确定了核多角蛋白基因的大部分序列存在于BamHI-E及PstI-C片段上,其大小分别为7.8Kb和6.7Kb.PstI-C片段经BamHI酶解后出现大小为4.0Kb和2.610的两个片段,将这两个片段克隆于Bluescript质粒中,并分析了4.0Kb片段其PstI端的部分DNA序列。
Viral DNA was extracted from the nuclear polyhedrosis virus (NPV) of the Chinese oak silkworm (Antheraea perxyz) and digested with restriction endonucleases PstI and BamHI; 31 and 6 restriction fragments were obtained, respectively.It was determined that the polyhedrin gene was located on the BamHI-E and PstI-C restriction fragments of ApNPV DNA by Southern hybridization with AcNPV polyhedrin gene fragment (HindIII- V restriction fragment of AcNPV) as DNA probe.The sizes of BamHI-C and PstI-C restriction fragments were 7. 8 kb and 6. 7 kb, respectively.After digestion with BamHI. the 6. 7 kb PstI-C restriction fragment produced two fragments of 4. 0 and 2. 7 kb. The two restriction fragment were recombined with Bluescript plasmid and used to transform E. coli DH5. One of the resulting recombined plasmids containing the 4. 0 kb restriction fragment was sequenced. a 124 bp DNA sequence of PstI-C restriction fragment was determined.
出处
《华南农业大学学报》
CAS
CSCD
1990年第4期35-39,共5页
Journal of South China Agricultural University
关键词
柞蚕
核型多角体
病毒
基因工程
Chinese oak silkworm
Antheraea pernyi
Nuclear polyhedrosis virus
gene cloning
