Expression of human soluble TRAIL in Chlamydomonas reinhardtii chloroplast

文献类型： 外文期刊

第一作者： Yang, Zongqi

作者： Yang, Zongqi;Li, Yinu;Chen, Feng;Li, Dong;Zhang, Zhifang;Liu, Yanxin;Zheng, Dexian;Wang, Yong;Shen, Guifang

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关键词： TRAIL (TNF-related apoptosis-inducing ligand);C. reinhardtii chloroplast;chloroplast expression;homologous recom-binant

期刊名称：CHINESE SCIENCE BULLETIN （ 影响因子：1.649； 五年影响因子：1.738 ）

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收录情况： SCI

摘要： Tumor necrosis factor-related apop-tosis-inducing ligand (TRAIL) induces selectively apoptosis in various tumor cells and virus-infected cells, but rarely in normal cells. A chloroplast expression vector, p64TRAIL, containing the cDNA coding for the soluble TRAIL (sTRAIL), was constructed with clpP-trnL-petB-chlL-rpl23-rpl2 as Chlamydomonas reinhardtii plastid homologous recombinant fragments and spectinomycin-resistant aadA gene as a select marker. The plasmid p64TRAIL was transferred into the chloroplast genome of C. reinhardtii by the biolistic method. Three independently transformed lines were obtained by 100 mg/L spectino-mycin selection. PCR amplification, Southern blot analysis of the sTRAIL coding region DNA and cultivation cells in the dark all showed that the exogenous DNA had been integrated into chloroplast genome of C. reinhardtii. Western blot analysis showed that human soluble TRAIL was expressed in C. reinhardtii chloroplast. The densitometric analysis of Western blot indicated that the expressed human sTRAIL protein in the chloroplasts of C. reinhardtii accounted for about 0.43 percent-0.67 percent of the total soluble proteins. These experimental results demonstrated the possibility of using transgenic chloroplasts of green alga as bioreactors for production of biopharmaceuticals.

分类号： N1

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