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编码LIM结构域蛋白CRP2的人类CSRP2基因的克隆、结构分析及染色体定位。

Cloning, structural analysis, and chromosomal localization of the human CSRP2 gene encoding the LIM domain protein CRP2.

作者信息

Weiskirchen R, Erdel M, Utermann G, Bister K

机构信息

Institute of Biochemistry, University of Innsbruck, Austria.

出版信息

Genomics. 1997 Aug 15;44(1):83-93. doi: 10.1006/geno.1997.4855.

Abstract

The CSRP2 gene encoding the LIM domain protein CRP2 was originally identified in quail based on its strong transcriptional suppression in transformed avian fibroblasts. Here we have isolated a human CSRP2 cDNA clone encoding a 193-amino-acid human CRP2 (hCRP2) protein with 96.4% amino acid sequence identity to the avian homolog. The CSRP2 cDNA clone was used to isolate CSRP2-related clones from gamma EMBL3 and P1 libraries of human genomic DNA. The complete organization of the CSRP2 gene was determined by nucleic acid hybridization, transcriptional mapping, and nucleotide sequence analysis. The gene spans a total of approximately 22 kb and contains six exons. The coding region is confined to exons 2-6 and predicts a hCRP2 protein identical in its amino acid sequence to the protein deduced from the CSRP2 cDNA clone. By fluorescence in situ hybridization using both lambda EMBL3 and P1 library clones as hybridization probes and a new method for computerized signal localization, CSRP2 was mapped to chromosome subband 12q21.1, a region frequently affected by deletion or breakage events in various tumor types. The library screens also led to the isolation of a CSRP2-related pseudogene, CSRP2P, which carried several extensive deletions and nucleotide substitutions but no intervening sequences in comparison to the CSRP2 cDNA sequence. By physical linkage and fluorescence in situ hybridization, CSRP2P was mapped to chromosome subband 3q21.1.

摘要

编码LIM结构域蛋白CRP2的CSRP2基因最初是在鹌鹑中发现的,基于其在转化的禽类成纤维细胞中的强烈转录抑制作用。在此,我们分离出了一个人类CSRP2 cDNA克隆,它编码一种由193个氨基酸组成的人类CRP2(hCRP2)蛋白,与禽类同源物的氨基酸序列同一性为96.4%。该CSRP2 cDNA克隆被用于从人类基因组DNA的γ EMBL3和P1文库中分离CSRP2相关克隆。通过核酸杂交、转录图谱分析和核苷酸序列分析确定了CSRP2基因的完整结构。该基因全长约22 kb,包含6个外显子。编码区局限于外显子2至6,预测的hCRP2蛋白的氨基酸序列与从CSRP2 cDNA克隆推导的蛋白相同。通过使用λ EMBL3和P1文库克隆作为杂交探针的荧光原位杂交以及一种新的计算机信号定位方法,将CSRP2定位到染色体亚带12q21.1,该区域在各种肿瘤类型中经常受到缺失或断裂事件的影响。文库筛选还导致分离出一个CSRP2相关的假基因CSRP2P,与CSRP2 cDNA序列相比,它有几个广泛的缺失和核苷酸取代,但没有间隔序列。通过物理连锁和荧光原位杂交,将CSRP2P定位到染色体亚带3q21.1。

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