摘要
应用酸性胍一步法从分泌 TLSF_(JM)的细胞系 JM 中提取总 RNA,进而纯化 mRNA,并应用微注射技术将纯化的 mRNA 注入非洲爪蛙卵母细胞中进行翻译表达。结果表明:非洲爪蛙卵母细胞能有效地翻译外源注入的 mRNA,翻译的 TLSF_(JM)产物主要滞留在卵母细胞内,仅有少量分泌到卵母细胞培养液中.TLSF_(JM) mRNA 体外翻译的成功为其 CDNA 文库的构建以及 TLSF_(JM)基因克隆和表达提供了重要的理论依据和实验基础.
Total RNA from JM cell lines secreting T leukmia derived suppressor factor was isolated using single—step method of acid guanidium followed by oligo—d(T)—cellulose affinity chromotogra-phy for purification of mRNA.TLSF_(JM)mRNA was mieroinjected into Xenopus Laevis Oocytes.The inhibitory activity in Xenopus Laevis Oocytes culture supernatant or lysate was detected by PHA plus IL—2 induced—mouse thymocyte proliferation assay The results showed that Xeno-pus Laevis Oocytes could efficiently translate exogenous mRNA,the trans ated products mainly existed inside oocytes and approximately2% of the products was secreted into the culture super-natant.Success in TLSF_(JM)mRNA translation in vitro might provide the basis for establishment of cDNA library and TLSF_(JM)gene cloning and expression.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
1992年第4期206-208,共3页
Chinese Journal of Immunology
基金
国家自然科学基金
关键词
抑制因子
RNA
微注射
体外翻译
Inhibitory factor
RNA
Microinjection
Translation in vitro
