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首页> 外文期刊>Molecular and Cellular Biology >Characterization of the gene and mRNA of the large subunit of ribulose-1,5-bisphosphate carboxylase in pea plants.
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Characterization of the gene and mRNA of the large subunit of ribulose-1,5-bisphosphate carboxylase in pea plants.

机译：豌豆植物中丝纤维 - 1,5-双磷酸羧酶大亚基基因及mRNA的表征。

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摘要

mRNA coding for the large subunit (LS) of ribulose-1,5-bisphosphate carboxylase was obtained by fractionating chloroplast polysomes on an affinity column, using anti-ribulose-1,5-bisphosphate carboxylase immunoglobulin G. Approximately 20% of the polysomal RNA specifically bound to the affinity column. LS mRNA was also isolated by fractionating chloroplast polysomal RNA on sucrose gradients. The LS mRNA fraction was identified by translation in vitro followed by immunoprecipitation with anti-ribulose-1,5-bisphosphate carboxylase immunoglobulin G. Labeled LS mRNA was hybridized to a genomic digests of pea chloroplast DNA. The LS gene was localized on a 3.55-kilobase pair BamHI fragment in SalI-SmaI DNA fragment 4. The BamHI fragment containing the LS gene was cloned, and a restriction endonuclease map was constructed. The LS gene was localized on a 1.9-kbp KpnI-EcoRI fragment. The LS gene was analyzed by electron microscopy, using the R loop mapping technique. LS mRNA was colinear with the gene, and its size was 1.35 +/- 0.2 kilobase pairs. When the LS mRNA was analyzed on methylmercury agarose gels, it comigrated with the 16S rRNA. The direction of transcription of the LS gene was in the same direction as that of the rRNA genes.
机译：通过在亲和柱上分离叶绿体 - 1,5-双磷酸羧化酶免疫球蛋白G.约20％的多晕染液相色谱法通过分离叶绿体 - 1,5-双磷酸碱基羧化酶的MRNA编码核苷酸-1,5-双磷酸羧化酶。特别绑定到亲和柱。通过在蔗糖梯度上分离叶绿体多晕RNA来分离LS mRNA。通过在体外翻译鉴定LS mRNA部分，然后用抗核糖糖糖-1,5-双磷酸羧化酶免疫球蛋白G.标记的LS mRNA与豌豆叶状物体DNA的基因组消化杂交的免疫沉淀。 LS基因在Sali-SmaI DNA片段中的3.55千碱基对BamHI片段中定位。克隆含有LS基因的BamHI片段，构建限制性内切核酸酶图。 LS基因在1.9kbp kpni-EcoRi片段上定位。通过电子显微镜通过R循环映射技术分析LS基因。 LS mRNA是基因的CLINEAR，其尺寸为1.35 +/- 0.2千碱基对。当在甲基汞琼脂糖凝胶上分析LS mRNA时，它用16S rRNA调用。 LS基因的转录方向与RRNA基因的转向相同。

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