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人血小板生成素氨基端功能区cDNA的克隆及其定点突变
引用本文:吴清发 桂向东. 人血小板生成素氨基端功能区cDNA的克隆及其定点突变[J]. 生物技术, 1998, 8(5): 10-12
作者姓名:吴清发 桂向东
作者单位:安徽省生物工程中心实验室!合肥230031
摘    要:
以Nested-PCR方法从人肝cDNA基因文库中扩增出编码人血小板生成素(hTPO)前153个氨基酸的氨基端功能区cDNA;在扩增中,采用非连续多核甘酸定点突变的方法.将翻译起始的七个氨基酸的原核中不常用的密码子同又突变成使用频率较高的密码子,以便于其在大肠杆菌中表达。序列测定证实了预期的结果。

关 键 词:人血小板成素  Nested-PCR  cDNA  定点突变  序列测定

AMINO-TERMINAL FUNCTIONAL REGION cDNA SYNTHSIS, CLONING AND SITE-DIRECTED MUTAGENESIS OF hTPO
Wu Qingfa, Gui Xiangdong, Fan Qinglin, Wei Yuewu, Song Lihua. AMINO-TERMINAL FUNCTIONAL REGION cDNA SYNTHSIS, CLONING AND SITE-DIRECTED MUTAGENESIS OF hTPO[J]. Biotechnology, 1998, 8(5): 10-12
Authors:Wu Qingfa   Gui Xiangdong   Fan Qinglin   Wei Yuewu   Song Lihua
Abstract:
The cDNA. coding human thromhopoietin (hTPO ) amino - terminal functional region 153 amino acids, was obtained by Nested -PCR from human liver cDNA library. Using the synthetic primers, seven genetic codes was replaced by the synonmous codes used more frequently in E, coli during Nested -PCR, The sequencing confirmed the expected result.
Keywords:human thrombopoietin (hTPO)   Nested-PCR   cDNA site-directed mutagenesis   DNA sequencing
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