The ovalbumin gene. Insertion of ovalbumin gene sequences in chimeric bacterial plasmids.
The ovalbumin gene. Insertion of ovalbumin gene sequences in chimeric bacterial plasmids.
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卵清蛋白基因。
DOI：
10.1016/s0021-9258(18)71832-2
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发表时间：
1977
期刊：
影响因子：
--
通讯作者：
B. O’Malley
中科院分区：
文献类型：
--
作者：
L. McReynolds;J. Monahan;D. W. Bendure;S. Woo;G. Paddock;W. Salser;J. Dorson;R. Moses;B. O’Malley
通过克隆细菌转化子，扩增和纯化了双链卵清蛋白DNA。利用大肠杆菌DNA聚合酶I和初始转录物的自引能力，从卵清蛋白mRNA的完整互补DNA转录物合成双链DNA。经S.核酸酶处理后，用末端脱氧核苷酸转移酶将poly（dA）添加到3 ‘末端，将卵清蛋白基因与含有3 ’ -poly（dT）末端的线性质粒pMB9杂交。该杂交分子被用于转化大肠杆菌菌株X1849。通过与全长cDNA杂交，克隆的转化子含有30 - 53%的卵清蛋白DNA。用限制性内切酶Hha i对所分离的质粒进行处理，证实了插入片段的长度。琼脂糖凝胶电泳分离表明，在所测试的克隆中插入的DNA数量在680到1090个碱基对之间。
Double-stranded ovalbumin DNA was amplified and purified by the cloning of bacterial transformants. The double-stranded DNA was synthesized from a complete complementary DNA transcript of ovalbumin mRNA using Escherichia coli DNA polymerase I and the self-priming ability of the initial transcript. After S. nuclease treatment, poly(dA) was added to the 3‘ termini with terminal deoxynucleotidyltransferase and the ovalbumin gene was hybridized to a linear plasmid DNA, pMB9, containing 3‘-poly(dT) termini. This hybrid molecule was used to transform the E. coli strain X1849. The cloned transformants contained from 30 to 53% of the complete ovalbumin DNA as determined by hybridization with full length cDNA. The length of the inserts was confirmed by treatment of the isolated plasmids with the restriction enzyme Hha I. Separation of the fragments by agarose gel electrophoresis showed that the amount of inserted DNA in clones tested varied from 680 to 1090 base pairs.