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小鼠基因组中肌动蛋白相关序列的数量与组织

Number and organization of actin-related sequences in the mouse genome.

作者信息

Minty A J, Alonso S, Guénet J L, Buckingham M E

出版信息

J Mol Biol. 1983 Jun 15;167(1):77-101. doi: 10.1016/s0022-2836(83)80035-7.

Abstract

Recombinant plasmids containing cDNA sequences complementary to the two mouse striated-muscle actin messenger RNAs (pAF81, pAM91) and to a non-muscle actin mRNA (pAL41) have been used to examine the number and organization of actin-related sequences in the mouse genome. A large number (greater than 20) of actin-related sequences are detected on Southern blots of restricted mouse DNA, the majority of which hybridize to both the 5' and 3' ends of the actin-coding sequence, even under conditions revealing only sequences greater than 80% homologous to the actin cDNA probes. More stringent washing of these blots indicates that the two striated muscle actins are each encoded by single genes, and that a non-muscle (beta or gamma) actin cDNA detects one homologous and two closely related sequences in mouse DNA. The segregation of the two striated-muscle actin genes in recombinant inbred mouse strains shows that these genes are not closely linked (greater than 1 centimorgan), and that the skeletal muscle actin gene is not linked to a non-muscle actin gene. Screening a bank of mouse genomic DNA, cloned in Charon 4A, indicates that the number of actin-related sequences in the mouse genome is much higher than 20. In particular, five phages have been isolated representing part of a sub-family of 20 to 50 similar but non-identical sequences, only weakly homologous to actin cDNA probes (probably a family of actin pseudogenes), which are the result of a recent amplification of a greater than 17 X 10(3) base region of mouse DNA.

摘要

含有与两种小鼠横纹肌肌动蛋白信使核糖核酸(pAF81、pAM91)以及一种非肌肉肌动蛋白信使核糖核酸(pAL41)互补的cDNA序列的重组质粒,已被用于检测小鼠基因组中肌动蛋白相关序列的数量和组织方式。在限制性内切酶消化的小鼠DNA的Southern印迹上检测到大量(超过20个)肌动蛋白相关序列,其中大多数与肌动蛋白编码序列的5'端和3'端均杂交,即使在仅显示与肌动蛋白cDNA探针同源性大于80%的序列的条件下也是如此。对这些印迹进行更严格的洗涤表明,两种横纹肌肌动蛋白各自由单个基因编码,并且一种非肌肉(β或γ)肌动蛋白cDNA在小鼠DNA中检测到一个同源序列和两个密切相关的序列。两种横纹肌肌动蛋白基因在重组近交小鼠品系中的分离表明,这些基因并非紧密连锁(大于1厘摩),并且骨骼肌肌动蛋白基因与非肌肉肌动蛋白基因不连锁。对克隆于Charon 4A中的小鼠基因组DNA文库进行筛选表明,小鼠基因组中肌动蛋白相关序列的数量远高于20个。特别地,已分离出五个噬菌体,它们代表了一个由20至50个相似但不相同的序列组成的亚家族的一部分,这些序列与肌动蛋白cDNA探针仅有微弱的同源性(可能是一个肌动蛋白假基因家族),它们是小鼠DNA中一个大于17×10³碱基区域近期扩增的结果。

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