腺病毒和腺伴随病毒基因元件提高白细胞介素6在真核细胞中的表达效率

STUDIES ON THE GENE ELEMENTS OF ADENOVIRUS AND ADENO ASSOCIATED VIRUS TO INCREASE THE EXPRESSION EFFICIENCY OF RECOMBINANT HUMAN IL 6 IN EUKARYOTIC CELLS

目的：构建高效真核表达载体以提高细胞因子在真核细胞中的表达效率。方法：利用基因克隆技术构建了两个新的白细胞介素６（ＩＬ６）真核表达载体ｐＡｄＣＩＩＬ６和ｐ３３５ＣＩＬ６。将它们瞬时转染ＣＯＳ７细胞后，利用ＩＬ６依赖株７ＴＤ１对上清中的ＩＬ６进行检测。结果：两种新建载体分别能提高ＩＬ６在真核细胞中的表达效率５．５和３．４倍。结论：腺病毒的病毒相关Ⅰ和ⅡＲＮＡ（ｖｉｒｕｓａｓｏｃｉａｔｅｄⅠａｎｄⅡＲＮＡ，ＶＡⅠａｎｄＶＡⅡ）和腺伴随病毒（ａｄｅｎｏａｓｏｃｉａｔｅｄｖｉｒｕｓ，ＡＡＶ）的倒转末端重复序列（ｉｎｖｅｒｔｅｄｔｅｒｍｉｎａｌｒｅｐｅａｔ。

Objective: To improve the expression efficiency of recombinant IL 6 in eukaryotic cells. Methods: Two new vectors (pAdCIIL 6, p335CIIL 6) were constructed by gene clone techniques. The bioactivity of IL 6 in culture supernatants of COS 7 cell transiently transfected by the new vectors was detected with IL 6 dependent cell line 7TD1 by MTT method. Results: The new vectors pAdCIIL 6 and p335CIIL 6 could enhance the expression efficiency of IL 6 in COS 7 cell 5.5 or 3.4 times higher than that of pcDIIL 6, respectively. Conclusion: Virus associated Ⅰ and Ⅱ RNA of adenovirus and inverted terminal repeat of adeno associated virus could enhance the expression efficiency of cytokine in eukaryotic cells. Construction of vectors with high expression efficiency is beneficial to in vivo function study of cytokine and gene therapy.