重组人干扰素α1b空间诱变菌种初步筛选研究

王俊锋; 刘金毅; 方向群; 徐晨; 郭英华; 刘桂东; 常德; 陆小冬; 刘长庭

doi:10.3969/j.issn.2095-5227.2013.01.005

重组人干扰素α1b空间诱变菌种初步筛选研究

Preliminary screening for space mutant strains of recombinant human interferonα1b

摘要

摘要: 目的利用“神舟八号”飞船对表达重组人干扰素α1b的基因工程菌株进行空间诱变，通过菌种筛选，获得高表达目的产物的菌株。方法对重组人干扰素α1b空间搭载菌种进行平板分离、抗性平板初筛和试管复筛，进行诱导表达，通过2轮SDS-PAGE分析获取高产菌株。结果首先从300个克隆中筛出221株阳性克隆，再从194个表达重组人干扰素α1b单克隆中筛选到144株具有氨苄青霉素抗性的克隆；通过等量接种和小量试管表达，最终筛选到6株目的蛋白表达量和表达比例较高的克隆。结论通过初步筛选，获得6株空间诱变后较高表达的重组人干扰素α1b生物工程菌菌株。

Abstract: Objective To obtain the highly expressed space mutant strains by screening recombinant human interferonα1b in Shenzhou 8 spacecraft. Methods The space mutant strains of recombinant human interferonα1b were isolated with flat-panel and screened with resistant flat-panel and test-tube. Expression of recombinant human interferonα1b was induced and space mutant strains were obtained by SDS-PAGE analysis. Results Two hundred and twenty-one clones were screened from 300 positive clones.Then 144 ampicillin-resistant clones were screened from 194 monoclones expressing recombinant human interferonα1b. At last, six clones with a high expression level of recombinant human interferon α1b were screened after the same amount was inoculated and expressed in test-tube. Conclusion Six bioengineering space mutant strains with a high expression level of recombinant human interferon α1b can be obtained by primary screening.

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