首页> 外国专利> Expressions vectors containing Lambda P.sub.L promoter and T.sub. 1 T.sub. 2 RNA termination sequence, plasmids containing the vectors, hosts containing the plasmids and relatedmethods

Expressions vectors containing Lambda P.sub.L promoter and T.sub. 1 T.sub. 2 RNA termination sequence, plasmids containing the vectors, hosts containing the plasmids and relatedmethods

机译:含有Lambda P.sub.L启动子和T.sub.p.的表达载体1吨2 RNA终止序列,包含载体的质粒,包含质粒的宿主和相关方法

摘要

An improved vector upon introduction into a suitable host containing the thermolabile repressor C.sub.I renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site; a first restriction enzyme site permitting replacement of the ribosomal binding site which follows thereafter; a ribosomal binding site; an ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzyme site for inserting the gene in phase with the ATG codon; a T.sub.1 T.sub.2 rRNA transcription termination sequence; an origin of replication and a gene associated with a selectable or identificable phenotypic trait manifested when the vector is present in the host. The distance between the 3' end of the P.sub.L O.sub.L promoter and operator sequence and the 5' end of the N utilization site is less than about 80 base pairs and the distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs.PPPlasmids have been constructed from the vectors and used to produce bovine, chicken and porcine growth hormones, human apolipoprotein E and human superoxide dismutase.
机译:引入含有热不稳定性阻遏物C1的合适宿主中的改进载体使宿主能够实现所需基因的表达。载体是双链DNA分子,其以5'至3'顺序包括以下:来自λ噬菌体的启动子和操纵子PLOL; N利用地点;第一限制性酶切位点,其允许随后取代核糖体结合位点;核糖体结合位点; ATG起始密码子或DNA,其在将所需基因插入载体后转化为ATG起始密码子;第二个限制性酶切位点,用于插入与ATG密码子同相的基因; T.sub.1 T.sub.2 rRNA转录终止序列;当载体存在于宿主中时,复制起点和与选择性或可识别表型性状相关的基因就会出现。 P.L.O.L.启动子和操纵子序列的3'端与N利用位点的5'端之间的距离小于约80个碱基对,并且该启动子和3'端的3'端之间的距离小于约80个碱基对。 N利用位点和核糖体结合位点的5'端少于约300个碱基对。质粒已由载体构建,用于产生牛,鸡和猪的生长激素,人载脂蛋白E和人超氧化物歧化酶。

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