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非洲爪蟾白蛋白基因的分离与鉴定:文库扩增导致74K白蛋白基因序列丢失。

Isolation and characterisation of the Xenopus laevis albumin genes: loss of 74K albumin gene sequences by library amplification.

作者信息

May F E, Weber R, Westley B R

出版信息

Nucleic Acids Res. 1982 May 11;10(9):2791-807. doi: 10.1093/nar/10.9.2791.

Abstract

The blood of the frog X.laevis contains 2 albumins of 68,000 and 74,000 daltons which are encoded in the liver by two related mRNAs. When an amplified X.laevis DNA library was screened with cloned albumin cDNA only 68,000 dalton albumin gene sequences were isolated. Hybridisation of the albumin cDNA to Southern-blots of Eco R1 digested X.laevis DNA showed that the sequences present in the recombinants did not account for all the fragments which hybridised on the Southern-blots. This indicated that 74K albumin gene sequences exist but that they are not present in the amplified DNA library. A X.laevis genomic library was therefore constructed and screened for albumin genes without amplification. Both 68K and 74K albumin gene sequences were isolated. Recombinants containing 74K albumin gene sequences grew extremely poorly and this probably explains why the 74K albumin sequences were ot isolated from the amplified library. Characterisation of the cloned DNA indicates that there is one sequence coding for the 68K albumin but two different sequences coding for the 75K albumin.

摘要

非洲爪蟾(X.laevis)的血液中含有两种白蛋白,分子量分别为68,000道尔顿和74,000道尔顿,它们由肝脏中的两种相关mRNA编码。当用克隆的白蛋白cDNA筛选扩增的非洲爪蟾DNA文库时,仅分离出了68,000道尔顿白蛋白的基因序列。白蛋白cDNA与经Eco R1消化的非洲爪蟾DNA的Southern印迹杂交显示,重组体中存在的序列并不能解释Southern印迹上所有杂交的片段。这表明74K白蛋白基因序列存在,但在扩增的DNA文库中不存在。因此构建了一个非洲爪蟾基因组文库,并在不进行扩增的情况下筛选白蛋白基因。68K和74K白蛋白基因序列均被分离出来。含有74K白蛋白基因序列的重组体生长极其缓慢,这可能解释了为什么74K白蛋白序列未从扩增文库中分离出来。对克隆DNA的表征表明,有一个序列编码68K白蛋白,但有两个不同的序列编码75K白蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c308/320656/11d244a6a043/nar00378-0038-a.jpg

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