Cloning system for Candida glabrata using elements from the metallothionein-IIa-encoding gene that confer autonomous replication.
Cloning system for Candida glabrata using elements from the metallothionein-IIa-encoding gene that confer autonomous replication.
复制标题
使用来自金属硫蛋白-IIa 编码基因的元件的光滑念珠菌克隆系统，该基因赋予自主复制能力。
DOI：
10.1016/0378-1119(92)90678-i
复制
发表时间：
1992
期刊：
影响因子：
3.5
通讯作者：
Winge,DR
中科院分区：
文献类型：
--
作者：
Mehra,RK;Thorvaldsen,JL;Macreadie,IG;Winge,DR
酵母假丝酵母有两个不同的家族编码金属硫蛋白（mt）。其中一个基因座，在许多野生型菌株中表现出选择性和串联扩增。glabrata。本文证明了扩增的mt - iiagene含有自主复制序列（ARS）。这些元素已经被用来构建能够复制inC. glabrata的载体。theMT-IIagene中的arelement (s)定位在theMT-IIagene编码序列下游457 bp的片段上。虽然含有这个片段的质粒转化。当使用整个1.25 kbmt - iagene时，高频率的glabrata提高了转化子的稳定性和质粒的拷贝数。转换离岸金融中心。携带2μ circleARSofSaccharomyces的质粒可以形成微菌落，这表明2μ质粒的arelement只在有限的范围内复制。相反，携带3个mmt - iiagene拷贝的aC. glabratapplasmid能够进行转化。酵母。
The yeastCandida glabrataharbors two distinct families that encode metallothioneins (MTs). One of these loci, theMT-IIalocus, exhibits selective and tandem amplication in many wild type strains ofC. glabrata. The present paper demonstrates that the amplifiedMT-IIagene contains autonomously replicating sequences (ARS). TheseARSelements have been used to construct vectors capable of replicating inC. glabrata. TheARSelement(s) in theMT-IIagene were localized to a 457-bp segment downstream from theMT-IIacoding sequence. Although plasmids containing this fragment transformC. glabratawith high frequency, the stability of the transformants and the copy number of the plasmid improve when the entire 1.25-kbMT-IIagene is used. Transformation ofC. glabratawith plasmids carrying the 2μ circleARSofSaccharomyces cerevisiaeled to the formation of micro-colonies, indicating that theARSelements of 2μ plasmids replicate only to a limited extent inC. glabrata. Conversely, aC. glabrataplasmid carrying three copies of theMT-IIagene was able to transformS. cerevisiae.