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成年和胚胎期鸡珠蛋白基因在染色体DNA中的分析。

Analysis of the adult and embryonic chicken globin genes in chromosomal DNA.

作者信息

Engel J D, Dodgson J B

出版信息

J Biol Chem. 1978 Nov 25;253(22):8239-46.

PMID:711749
Abstract

Globin-specific mRNA has been purified from the red blood cells of anemic adult hens and from those of 5-day and 7-day chick embryos. These RNA preparations were used to synthesize labeled cDNA for solution and filter hybridization (Southern blotting) experiments. Hybridization of cDNAs prepared with separated adult alpha- and beta-mRNAs and of rabbit globin-specific probes to restriction enzyme-digested, filter-bound chicken chromosomal DNA has shown that the adult globin genes of the chicken exist within EcoRI-cleaved chromosomal DNA fragments which are 4.3, 6.0, and 12.5 kilobase pairs (kbp) in size. These fragments contain the majority or entirety of the adult alpha D, beta, and alpha A genes, respectively. Furthermore, we present evidence that the predominant chicken embryonic globins are contained within EcoRI fragments 9.4 and 15 kbp in size. Evidence is also presented that the same globin organization exists in several different breeds of chickens and in several chicken culture cell types. Finally, it is demonstrated that the pattern of globin-specific hybridization to reticulocyte chromosomal DNA digested with several restriction enzymes is unchanged during erythropoietic development.

摘要

已从成年贫血母鸡以及5日龄和7日龄鸡胚的红细胞中纯化出球蛋白特异性mRNA。这些RNA制剂用于合成标记的cDNA,以进行溶液杂交和滤膜杂交(Southern印迹)实验。用分离的成年α和β mRNA制备的cDNA以及兔球蛋白特异性探针与经限制性内切酶消化、固定在滤膜上的鸡染色体DNA杂交,结果表明,鸡的成年球蛋白基因存在于大小分别为4.3、6.0和12.5千碱基对(kbp)的EcoRI切割的染色体DNA片段中。这些片段分别包含成年α D、β和α A基因的大部分或全部。此外,我们提供的证据表明,主要的鸡胚胎球蛋白包含在大小为9.4和15 kbp的EcoRI片段中。还有证据表明,在几种不同品种的鸡和几种鸡培养细胞类型中存在相同的球蛋白组织。最后,证明了在用几种限制性酶消化的网织红细胞染色体DNA上,球蛋白特异性杂交模式在红细胞生成发育过程中保持不变。

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