Arnold H H, Siddiqui M A
Biochemistry. 1979 Dec 11;18(25):5641-7. doi: 10.1021/bi00592a019.
Double-stranded complementary deoxyribonucleic acid (cDNA) transcribed in vitro from a partially pure myosin light-chain messenger ribonucleic acid (mRNA) of the chick embryonic heart was cloned in Escherichia coli strain chi 1776 by using the HindIII cleavage site in the plasmid pBR322. The insertion of essentially full length DNA was achieved by repeated selection of large-size cDNA transcripts. Of the 12 transformants that contained large-size DNA inserts, the clone pML10 insert was 950 base pairs in length, almost the same size as myosin light-chain mRNA (980 nucleosides). The clone pML10 was identified by hybridization with a highly pure cDNA probe and by hybrid-arrested translation assay. pML10 was further characterized by partial restriction enzyme mapping. The availability of a cloned DNA probe for myosin light-chain facilitates the analysis of the mechanism underlying the induction of cardiac muscle specific gene transcription in presumptive heart-forming cells of the chick blastoderm.
从鸡胚心脏部分纯化的肌球蛋白轻链信使核糖核酸（mRNA）体外转录的双链互补脱氧核糖核酸（cDNA），利用质粒pBR322中的HindIII切割位点克隆于大肠杆菌菌株chi 1776。通过反复选择大尺寸的cDNA转录本实现了基本全长DNA的插入。在12个含有大尺寸DNA插入片段的转化子中，克隆pML10插入片段长度为950个碱基对，几乎与肌球蛋白轻链mRNA（980个核苷）大小相同。通过与高纯度cDNA探针杂交以及杂交捕获翻译试验鉴定了克隆pML10。通过部分限制性酶切图谱分析对pML10进行了进一步表征。用于肌球蛋白轻链的克隆DNA探针的可得性有助于分析鸡胚盘预定心脏形成细胞中心肌特异性基因转录诱导的潜在机制。
