Total synthesis and expression of a gene for the alpha-subunit of bovine rod outer segment guanine nucleotide-binding protein (transducin).
Total synthesis and expression of a gene for the alpha-subunit of bovine rod outer segment guanine nucleotide-binding protein (transducin).
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牛杆外段鸟嘌呤核苷酸结合蛋白（转导蛋白）α亚基基因的全合成和表达。
DOI：
10.1093/nar/16.14.6361
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发表时间：
1988
影响因子：
14.9
通讯作者：
Khorana,HG
中科院分区：
文献类型：
--
作者：
Sakmar,TP;Khorana,HG
为了便于通过位点特异性诱变进行结构-功能研究，我们合成了牛棒外段（ROS）鸟嘌呤核苷酸结合蛋白（转导蛋白）α-亚基的基因。该基因编码天然氨基酸序列，并设计包含38个均匀间隔的独特限制性内切位点。这使得通过限制性片段替换在基因的任何部分发生突变成为可能。该基因长度为1076个碱基对。它是由44个人工合成的寡核苷酸在体外酶切连接成4个片段而构建的。利用腺病毒主要晚期启动子控制转录的载体，在猴肾细胞（COS-1）中表达了合成转导素基因和编码转导素的cDNA，表达水平相近。
To facilitate structure-function studies by site-specific mutagenesis, we have synthesized a gene for the α-subunit of the bovine rod outer segment (ROS) guanine nucleotide-binding protein (transducin). The gene codes for the native amino acid sequence and contains, by design, 38 unique restriction sites which are uniformly spaced. This enables mutagenesis in any part of the gene by restriction fragment replacement. The gene is 1076 base pairs in length. It was constructed from 44 synthetic oligonucleotides which were joined enzymaticallyin vitrointo four fragments which were cloned. The synthetic transducin gene and cDNA encoding transducin were expressed to similar levels in monkey kidney cells (COS-1) using a vector in which transcription was under the control of the adenovirus major late promoter.