ISOLATION OF ALGAL GENES BY FUNCTIONAL COMPLEMENTATION OF YEAST1
ISOLATION OF ALGAL GENES BY FUNCTIONAL COMPLEMENTATION OF YEAST1
复制标题
通过酵母的功能互补分离藻类基因1
DOI：
10.1046/j.1529-8817.2002.01022.x
复制
发表时间：
2002
影响因子：
2.9
通讯作者：
K. Apt
中科院分区：
文献类型：
--
作者：
J. C. Lippmeier;A. M. Brown;K. Apt
分离藻类cDNA，并通过酵母营养缺陷型的功能互补来表征。使用酿酒酵母表达载体pFL-61构建了两个cDNA文库，一个来自硅藻三角褐指藻Bohlin，另一个来自甲藻隐甲藻Cohnii Biecheler。这些文库用于两种酵母菌株中营养缺陷型标记的功能互补。由三角褐指藻文库补充的酵母色氨酸营养缺陷型含有编码与色氨酸合成相关的双结构域蛋白质的质粒，即吲哚-3-甘油磷酸合酶-N-（5′-磷酸核糖基）邻氨基苯甲酸异构酶。另一个cDNA来源于C. cohnii library拯救了S.从腺嘌呤生物合成的缺陷中获得。该cDNA编码磷酸核糖氨基咪唑-琥珀酰胺合成酶和磷酸核糖氨基咪唑羧化酶的融合体，其分别对应于酵母ade 1和ade 2基因。这些结果表明，异源功能互补可用于鉴定藻类基因，并可能提供优于其他基因发现方法。
Algal cDNAs were isolated and characterized by functional complementation of yeast auxotrophs. Two cDNA libraries, one derived from the diatom Phaeodactylum tricornutum Bohlin and the other from the dinoflagellate Crypthecodinium cohnii Biecheler, were constructed using the Saccharomyces cerevisiae expression vector pFL‐61. These libraries were used for functional complementation of auxotrophic markers in two yeast strains. Yeast tryptophan auxotrophs, complemented by the P. tricornutum library, contained a plasmid that encoded a two‐domain protein associated with tryptophan synthesis, indole‐3‐glycerol phosphate synthase‐N‐(5′‐phosphoribosyl) anthranilate isomerase. Another cDNA originating from the C. cohnii library rescued S. cervisiae from a defect in adenine biosynthesis. This cDNA encoded a fusion of phosphoribosylamidoimidazole‐succinocarboxamide synthetase and phosphoribosylaminoimidazole carboxylase, which correspond to the yeast ade1 and ade2 genes, respectively. These results demonstrate that heterologous functional complementation can be used to identify algal genes and may provide advantages over other gene discovery methods.
分离编码酿酒酵母 GCN2 eIF-2α 激酶的黑腹果蝇同源物的基因。
影响因子：
3.3
作者：
Olsen,DS;Jordan,B;Chen,D;Wek,RC;Cavener,DR
通讯作者：
Cavener,DR
基于 Tn3 转座子诱变的定向 DNA 测序策略：应用于酿酒酵母 I 号染色体上的 ADE1 基因座。
影响因子：
14.9
作者：
Davies,CJ;Hutchison3rd,CA
通讯作者：
Hutchison3rd,CA