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Cloning of chicken lysozyme structural gene sequences synthesized in vitro.

机译:体外合成的鸡溶菌酶结构基因序列的克隆。

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摘要

Double-stranded chicken lysozyme cDNA was synthesized from an oviduct mRNA fraction enriched for lysozyme mRNA. The ds-cDNA was inserted into the BamHI site of plasmid pBR322 using chemically synthesized DNA linker molecules containing the BamHI restriction endonuclease cleavage site. After bacterial transformation, colonies carrying lysozyme DNA were identified by hybridization with highly purified lysozyme cDNA. The 555 base pairs long cloned DNA fragment of one recombinant plasmid was isolated and characterized by restriction endonuclease digestion. The DNA sequence of selected parts of the inserted DNA is as predicted from the amino acid sequence of prelysozyme. The sequence data allows the unambiguous location of the coding region within lysozyme mRNA.
机译:从富含溶菌酶mRNA的输卵管mRNA级分合成了双链鸡溶菌酶cDNA。使用含有BamHI限制性核酸内切酶切割位点的化学合成的DNA接头分子,将ds-cDNA插入质粒pBR322的BamHI位点。细菌转化后,通过与高度纯化的溶菌酶cDNA杂交来鉴定带有溶菌酶DNA的菌落。分离了一种重组质粒的555个碱基对长克隆的DNA片段,并通过限制性内切酶消化进行了表征。如从溶菌酶的氨基酸序列所预测的那样,插入的DNA的选定部分的DNA序列。序列数据允许溶菌酶mRNA内的编码区明确定位。

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