摘要
昆虫核型多角体病毒(NPV)P10基因属于晚期基因,为强启动子所控制,又是病毒复制所非必需的基因。我们对前报的家蚕核型多角体病毒(BmNPV)P10基因,应用PCR技术进行ATG区定点突变,在ATG被突变的同时形成一个BglⅡ酶切位点,得到一个不含ATG的BmNPV P10基因启动子。将长为230bp的经突变后的BmNPV P10基因5’端(包括启动子所有特征)片段克隆进pMMTV·CAT质粒中,构建成一个CAT基因在BmNPV P10基因启动子控制下的pBmPl0·CAT瞬间表达质粒。该质粒通过转染进入经野生型BmNPV感染的BmN细胞中,CAT得以表达。证明BmNPV P10启动子是比较强的启动子,可以在BmN细胞表达外源基因,具备了作为表达载体启动子的特性。
Insect nuclear polyhedrosis virus ( INPV ) p10 gene is a verylate gene which is controlled by a strong promoter and is not essential for virus replication. According to our previous report concerning the cloning and sequencing of p10 gene from Bombyx mori nuclear polyhedrosis virus ( BmNPV), we designed a procedure so that the ATG region of p10 gene from BmNPV was mutanted by PCR, at the same time this region became a Bg1 Ⅱ site, obtaining a p10 promoter with no ATG start codon. The complete p10 promoter in a 230 bp fragment was cloned into pMMTV CAT. Thus, we obtained pBmp10·2CAT as a transient expression vector. The CAT gene was controlled by p10 promoter from BmNPV in the vector. pBmp10·CAT was transfected into BmNPV-infected BmN celis. The rusult indicates that BmNPV p10 promoter can express CAT gene at high Ievel in BmN celis. The p10 promoter of BmNPV is qualified as a hype-rexpression vector promoter.
出处
《病毒学报》
CAS
CSCD
北大核心
1993年第4期361-366,共6页
Chinese Journal of Virology
基金
国家八.五攻关项目
关键词
家蚕
核多角体病毒
基因
启动子
Bombyx mori
Nuclear polyhedrosis virus
P10 gene
Promoter
pCR
CAT gene
Expression
