ISOLATION AND CHARACTERIZATION OF HUMAN ACTIN GENES
ISOLATION AND CHARACTERIZATION OF HUMAN ACTIN GENES
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DOI：
10.1073/pnas.78.8.4674
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
KEDES, L
中科院分区：
文献类型：
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作者：
ENGEL, JN;GUNNING, PW;KEDES, L
利用来自果蝇和鸡的克隆肌动蛋白基因从人类 DNA 文库中分离出 12 个肌动蛋白基因片段。通过体外翻译测定，这 12 个克隆中的每一个均具有选择性与人肌动蛋白 mRNA 杂交的能力，显示其含有肌动蛋白编码区。根据在 1 维和二维十二烷基硫酸钠/聚丙烯酰胺凝胶上电泳时与真实肌动蛋白的共迁移以及根据其部分蛋白水解产物，将翻译产物判断为肌动蛋白。通过限制性内切核酸酶消化这些重组噬菌体中的每一个所产生的片段的大小和顺序的测定表明它们是非等位基因的并且来自基因组的非重叠区域。这些克隆的人类肌动蛋白基因以及果蝇和鸡肌动蛋白基因克隆用于表明人类基因组包含25-30个与肌动蛋白基因同源的EcoRI片段，并且在测试的3个非近亲个体中，这些片段均不表现出长度或多态性。
Cloned actin genes from Drosophila melanogaster and from chicken were utilized to isolate 12 actin gene fragments from a human DNA library. Each of these 12 clones was shown to contain actin coding regions by its ability to selectively hybridize to human actin mRNA as assayed by in vitro translation. The translation product was judged to be actin on the basis of its comigration with authentic actins when electrophoresed on 1- and 2-dimensional sodium dodecyl sulfate/polyacrylamide gels and on the basis of its partial proteolysis products. Determination of the sizes and order of the fragments generated by restriction endonuclease digestion of each of these recombinant phages shows that they are nonallelic and are from nonoverlapping regions of the genome. These cloned human actin genes and the Drosophila and chicken actin gene clones were utilized to show that the human genome contains 25-30 EcoRI fragments homologous to actin genes and that, among 3 nonconsanguineous individuals tested, none of these fragments exhibit length or restriction-site polymorphism.