Westhoff P, Zetsche K
Eur J Biochem. 1981 May 15;116(2):261-7. doi: 10.1111/j.1432-1033.1981.tb05328.x.
Poly(A)-rich and poly(A)-free RNAs were isolated from autotrophic and heterotrophic cells of the phytoflagellate Chlorogonium elongatum and translated in an mRNA-depleted reticulocyte lysate system. Immunoprecipitation methods were improved to detect large and small subunits of the chloroplast enzyme ribulose-1,5-bisphosphate carboxylase synthesized in vitro. Large-subunit polypeptides were shown to be the translation products of poly(A)-free RNA having the same molecular weight as large subunits made in vivo. Small-subunit polypeptides were synthesized when poly(A)-rich RNA was used as a template. They were made in vitro as a precursor, with an Mr about 6000 larger than mature small subunits. Cells growing heterotrophically in the dark with acetate are provided with lower levels of mRNA activities for the large and the small subunits is at least partially controlled by the amounts of translatable mRNAs.
从植物鞭毛虫长绿梭藻的自养和异养细胞中分离出富含多聚腺苷酸(Poly(A))和无多聚腺苷酸的RNA,并在无mRNA的网织红细胞裂解物系统中进行翻译。改进了免疫沉淀方法,以检测体外合成的叶绿体酶核酮糖-1,5-二磷酸羧化酶的大亚基和小亚基。大亚基多肽被证明是无多聚腺苷酸RNA的翻译产物,其分子量与体内合成的大亚基相同。当使用富含多聚腺苷酸的RNA作为模板时,会合成小亚基多肽。它们在体外以前体形式合成,其分子量比成熟小亚基大约6000。在黑暗中以乙酸盐为营养源进行异养生长的细胞,其大亚基和小亚基的mRNA活性水平较低,这至少部分受可翻译mRNA量的控制。