Nudel U, Katcoff D, Carmon Y, Zevin-Sonkin D, Levi Z, Shaul Y, Shani M, Yaffe D
Nucleic Acids Res. 1980 May 24;8(10):2133-46. doi: 10.1093/nar/8.10.2133.
The construction and identification of a recombinant plasmid containing a cDNA insert which hybridizes specifically to myosin heavy chain mRNA is described. The plasmid was used as a probe to screen a rat genomic library for recombinant phages containing myosin heavy chain sequences. Six clones with approximately 15 k bp inserts each were isolated. Digestion with several restriction enzymes and hybridization of the fractionated DNA with the plasmid probe showed that the clones contained 3 different DNA inserts. Electron microscopy of a heteroduplex made by hybridization of DNA from two clones confirmed that the inserts originated in different genes. Hybridization of size-fractionated ECOR1 digested rat spleen DNA with the cloned probe suggested the existence of at least 5 myosin heavy chain genes.
本文描述了一种重组质粒的构建与鉴定，该质粒含有一个能与肌球蛋白重链mRNA特异性杂交的cDNA插入片段。该质粒用作探针，筛选大鼠基因组文库以寻找含有肌球蛋白重链序列的重组噬菌体。分离出六个克隆，每个克隆含有约15kbp的插入片段。用几种限制性酶消化，并将分级分离的DNA与质粒探针杂交，结果表明这些克隆含有3种不同的DNA插入片段。对两个克隆的DNA杂交形成的异源双链体进行电子显微镜观察，证实插入片段起源于不同的基因。用克隆探针与经大小分级的EcoR1消化的大鼠脾脏DNA杂交，提示至少存在5个肌球蛋白重链基因。
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