CAJ | 학술논문

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.
목적이용“신주팔호”비선대표체중조인간우소α1b적기인공정균주진행공간유변,통과균충사선,획득고표체목적산물적균주.방법대중조인간우소α1b공간탑재균충진행평판분리、항성평판초사화시관복사,진행유도표체,통과2륜SDS-PAGE분석획취고산균주.결과수선종300개극륭중사출221주양성극륭,재종194개표체중조인간우소α1b단극륭중사선도144주구유안변청매소항성적극륭;통과등량접충화소량시관표체,최종사선도6주목적단백표체량화표체비례교고적극륭.결론통과초보사선,획득6주공간유변후교고표체적중조인간우소α1b생물공정균균주.
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 interferonα1b 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 interferonα1b can be obtained by primary screening.