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一个与人类酰基辅酶A结合蛋白/地西泮结合抑制剂相关的假基因的分子克隆及染色体定位

Molecular cloning and chromosomal localization of a pseudogene related to the human acyl-CoA binding protein/diazepam binding inhibitor.

作者信息

Gersuk V H, Rose T M, Todaro G J

机构信息

Virginia Mason Research Center, Department of Immunology, Seattle, Washington 98101, USA.

出版信息

Genomics. 1995 Jan 20;25(2):469-76. doi: 10.1016/0888-7543(95)80047-p.

Abstract

The acyl-CoA binding protein (ACBP) and the diazepam binding inhibitor (DBI) or endozepine are independent isolates of a single 86-amino-acid, 10-kDa protein. ACBP/DBI is highly conserved between species and has been identified in several diverse organisms, including human, cow, rat, frog, duck, insects, plants, and yeast. Although the genomic locus has not yet been cloned in humans, complementary DNA clones with different 5' ends have been isolated and characterized. These cDNA clones appear to be encoded by a single gene. However, Southern blot analyses, in situ hybridizations, and somatic cell hybrid chromosomal mapping all suggest that there are multiple ACBP/DBI-related sequences in the genome. To identify potential members of this gene family, degenerate oligonucleotides corresponding to highly conserved regions of ACBP/DBI were used to screen a human genomic DNA library using the polymerase chain reaction. A novel gene, DBIP1, that is closely related to ACBP/DBI but is clearly distinct was identified. DBIP1 bears extensive sequence homology to ACBP/DBI but lacks the introns predicted by rat and duck genomic sequence studies. A 1-base deletion in the coding region results in a frameshift and, along with the absence of introns and the lack of a detectable transcript, suggests that DBIP1 is a pseudogene. ACBP/DBI has previously been mapped to chromosome 2, although this was recently disputed, and a chromosome 6 location was suggested. We show that ACBP/DBI is correctly placed on chromosome 2 and that the gene identified on chromosome 6 is DBIP1.

摘要

酰基辅酶A结合蛋白(ACBP)与地西泮结合抑制剂(DBI)或内源性苯二氮䓬是一种由86个氨基酸组成、分子量为10 kDa的单一蛋白质的独立分离物。ACBP/DBI在物种间高度保守,已在多种不同生物中被鉴定出来,包括人类、牛、大鼠、青蛙、鸭子、昆虫、植物和酵母。尽管人类的基因组位点尚未被克隆,但已分离并鉴定了具有不同5'端的互补DNA克隆。这些cDNA克隆似乎由单个基因编码。然而,Southern印迹分析、原位杂交和体细胞杂交染色体定位均表明基因组中存在多个与ACBP/DBI相关的序列。为了鉴定该基因家族的潜在成员,使用对应于ACBP/DBI高度保守区域的简并寡核苷酸,通过聚合酶链反应筛选人类基因组DNA文库。鉴定出了一个与ACBP/DBI密切相关但明显不同的新基因DBIP1。DBIP1与ACBP/DBI具有广泛的序列同源性,但缺乏大鼠和鸭子基因组序列研究预测的内含子。编码区的一个单碱基缺失导致移码,加上缺乏内含子和未检测到转录本,表明DBIP1是一个假基因。ACBP/DBI先前已被定位到2号染色体,尽管最近对此存在争议,并有人提出其位于6号染色体上。我们表明ACBP/DBI正确地定位于2号染色体,而在6号染色体上鉴定出的基因是DBIP1。

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