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人类精子黏附分子基因SPAM1的表达分析、基因组结构及定位到7q31

Expression analysis, genomic structure, and mapping to 7q31 of the human sperm adhesion molecule gene SPAM1.

作者信息

Jones M H, Davey P M, Aplin H, Affara N A

机构信息

University of Cambridge Department of Pathology, United Kingdom.

出版信息

Genomics. 1995 Oct 10;29(3):796-800. doi: 10.1006/geno.1995.9931.

Abstract

During the course of systematic sequence tag analysis of clones isolated from an adult testis cDNA library, clones 296 and 576 were found to detect 71-74% sequence identity to the guinea pig sperm surface protein PH-20. This surface protein is involved in sperm-egg adhesion in the guinea pig. Nucleotide sequence for 1919 bp of human DNA from a series of overlapping cDNA clones isolated from a testis cDNA library confirmed the sequence identity within a 1527-bp open reading frame to be 71-74% to the guinea pig gene and the similarity to be 60% for the predicted protein of 509 amino acids. Southern blot analysis of human genomic DNA and DNA from somatic cell hybrids indicates that the gene (SPAM1) is unique and does not form part of a larger family and that it maps to chromosome 7. Fluorescence in situ hybridization with yeast artificial chromosome (YAC) clones isolated from the CEPH megaYAC library has refined this localization to 7q31. PCR analysis of genomic DNA and YAC clone DNA has shown that the 1919 bp of the gene that has been cloned covers approximately 11 kb of genomic DNA and is encoded by at least 4 exons. Northern analysis of poly(A)+ mRNA from a range of 16 human tissues has demonstrated that expression of the gene as a single 2.4-kb transcript is strictly limited to the testis.

摘要

在对从成人睾丸cDNA文库中分离出的克隆进行系统序列标签分析的过程中,发现克隆296和576与豚鼠精子表面蛋白PH-20的序列同一性为71-74%。这种表面蛋白参与豚鼠的精卵黏附。从睾丸cDNA文库中分离出的一系列重叠cDNA克隆对1919 bp的人类DNA进行核苷酸测序,证实了1527 bp开放阅读框内与豚鼠基因的序列同一性为71-74%,对预测的509个氨基酸的蛋白质的相似性为60%。对人类基因组DNA和体细胞杂种DNA的Southern印迹分析表明,该基因(SPAM1)是独特的,不构成更大基因家族的一部分,并且它定位于7号染色体。用从CEPH megaYAC文库中分离出的酵母人工染色体(YAC)克隆进行荧光原位杂交,已将该定位精确到7q31。对基因组DNA和YAC克隆DNA的PCR分析表明,已克隆的1919 bp基因覆盖了约11 kb的基因组DNA,由至少4个外显子编码。对来自16种人类组织的poly(A)+ mRNA进行Northern分析表明,该基因作为单一的2.4 kb转录本的表达严格限于睾丸。

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