| 大鼠α-酰胺酶在变铅青链霉菌中的克隆及表达研究 |
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| 引用本文： | 武兵元,李元.大鼠α-酰胺酶在变铅青链霉菌中的克隆及表达研究[J].生物工程学报,2000,16(5):574-577. |
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| 作者姓名： | 武兵元 李元 |
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| 作者单位： | 中国医学科学院院中国协和医科大学医药生物技术研究所,北？中国医学科学院院中国协和医科大学医药生物技术研 |
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| 基金项目： | 九五生物技术攻关项目经费资助(96-c02-01-08)。 |
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| 摘 要： | α－酰胺酶（α－ａｍｉｄａｓｅ，α－ＡＥ）催化神经和内分泌系统中活性多肽的Ｃ－端酰胺化，多多肽的生物活性至关重要。以大鼠心房组织的总ＲＮＡ为模板，采用ＲＴ－ＰＣＲ技术扩增获得编码α－酰胺酶的ｃＤＮＡ，并进行了克隆和测序。为了使α－酰胺酶能在链霉菌中分泌表达，将其ｃＤＮＡ与链霉菌酷氮酶酶（ｍｅｌＣ１）信号的编码序列融合得到融合ｍｅｌ／ＡＥ，将ｍｅｌ／ＡＥ插入链霉菌质粒ｐＩＪ６８０，获得重组质粒ｐＩＪ
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| 关 键 词： | 酰胺酶 基因克隆 变铅青链霉菌 酰胺化多肽 |
| 文章编号： | 1000-3061(2000)05-0574-04 |
| 修稿时间： | 1999-09-06 |
Studies of cloning and expression of rat alpha-amidase gene in Streptomyces lividans] |
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| WU Bin-Yuan
,LI Yuan.Studies of cloning and expression of rat alpha-amidase gene in Streptomyces lividans][J].Chinese Journal of Biotechnology,2000,16(5):574-577. |
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| Authors: | WU Bin-Yuan LI Yuan |
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| Institution: | WU Bin-Yuan
,LI Yuan
;(Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Peking Medical College, Beijing 100050) |
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| Abstract: | alpha-Amidase catalyzes the C-terminal amidation of active polypeptides in the Nerve-Endocrine system. It is important for full biological activity of the polypeptides. By using the total RNA of rat atrium as a templet, the cDNA encoding alpha-amidase was amplified by RT-PCR and sequenced. The cDNA was fused with the coding sequence of mel C1 signal peptide to produce fusion gene mel/AE for secretive expression in Streptomyces lividans. The fusion gene mel/AE was inserted into plasmid pIJ680 and the recombinant plasmid pIJ-mel/AE680 was obtained. The results of SDS-PAGE and biological activity showed that the recombinant strain S. lividans TK54pIJ-mel/AE 680] produced secretly alpha-amidase. |
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| Keywords: | amidase gene cloning and expression Streptomyces lividans |
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