一种新型高容量黏粒载体及其应用。

A new high-capacity cosmid vector and its use.

作者信息

Meyerowitz E M, Guild G M, Prestidge L S, Hogness D S

出版信息

Gene. 1980 Nov;11(3-4):271-82. doi: 10.1016/0378-1119(80)90067-0.

Abstract

The construction of a cosmid, MUA-3, designed for the convenient cloning of eukaryotic DNA segments up to 48 kb in length is described. The cosmid contains all of the plasmid pBR322 with approx. 400 bases of lambda DNA, including the cohesive end site, inserted at the pBR322 PstI endonuclease recognition site. Methods for using this vector to construct several types of Drosophila melanogaster genomic DNA libraries are given, and libraries made by these methods are characterized. A sheared Drosophila DNA-EcoRI linker library is shown to stably maintain average Drosophila DNA inserts of over 40 kb and up to 48 kb, and the efficiency of producing clones by a partial restriction and ligation method is shown to be over 3 X 10(5) clones/microgram of Drosophila DNA.

摘要

本文描述了粘粒MUA-3的构建，该粘粒旨在方便地克隆长度达48kb的真核DNA片段。该粘粒包含整个质粒pBR322以及约400个λDNA碱基，包括粘性末端位点，其插入到pBR322 PstI核酸内切酶识别位点。文中给出了使用该载体构建几种类型果蝇基因组DNA文库的方法，并对通过这些方法构建的文库进行了表征。一个经剪切的果蝇DNA-EcoRI接头文库被证明能稳定维持平均长度超过40kb且最长达48kb的果蝇DNA插入片段，并且通过部分酶切和连接方法产生克隆的效率被证明超过3×10⁵个克隆/微克果蝇DNA。