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秀丽隐杆线虫中精子特异性基因家族的分离与鉴定

Isolation and characterization of a sperm-specific gene family in the nematode Caenorhabditis elegans.

作者信息

Klass M R, Kinsley S, Lopez L C

出版信息

Mol Cell Biol. 1984 Mar;4(3):529-37. doi: 10.1128/mcb.4.3.529-537.1984.

Abstract

The major sperm protein (MSP) of the nematode Caenorhabditis elegans is a low-molecular-weight (15,000) basic protein implicated in the pseudopodial movement of mature spermatozoa. Its synthesis occurs in a specific region of the gonad and is regulated at the level of transcription (M. Klass and D. Hirsh, Dev. Biol. 84:299-312, 1981; S. Ward and M. Klass, Dev. Biol. 92:203-208, 1982; Klass et al., Dev. Biol. 93:152-164, 1982). A developmentally regulated gene family has been identified that codes for this MSP. Whole genomic blots, as well as analysis of genomic clone banks, indicate that there are between 15 and 25 copies of the MSP gene in the nematode genome. Southern blot analysis also indicates that there is no rearrangement or amplification within the MSP gene family during development. No evidence was found of methylation at various restriction sites surrounding the MSP gene family, and similarly, no correlation between methylation and expression was observed. Three distinct members of this MSP gene family have been cloned, and their nucleotide sequences have been determined. Differential screening of a cDNA clone bank made from polyadenylated mRNA from adult males yielded 45 male-specific clones, 32 of which were clones of MSP genes. One of these cDNA clones was found to contain the entire nucleotide sequence for the MSP, including part of the 5' leader and all of the 3' trailing sequence. Genomic clones bearing copies of the MSP genes have been isolated. At least one of the members of this gene family is a pseudogene. Another member of the MSP gene family that has been cloned from genomic DNA contains the entire uninterrupted structural sequence for the MSP in addition to a 5' flanking sequence containing a promoter-like region with the classic TATA box, a sequence resembling the CAAT box, and a putative ribosome-binding sequence. The 3' trailing sequences of the genomic and the cDNA clones contain an AATAAA polyadenylation site.

摘要

秀丽隐杆线虫的主要精子蛋白(MSP)是一种低分子量(15,000)的碱性蛋白,与成熟精子的伪足运动有关。它在性腺的特定区域合成,并在转录水平受到调控(M. 克拉斯和D. 赫什,《发育生物学》84:299 - 312,1981;S. 沃德和M. 克拉斯,《发育生物学》92:203 - 208,1982;克拉斯等人,《发育生物学》93:152 - 164,1982)。已鉴定出一个受发育调控的基因家族,该家族编码这种MSP。全基因组印迹以及基因组克隆文库分析表明,线虫基因组中MSP基因有15至25个拷贝。Southern印迹分析还表明,在发育过程中MSP基因家族内没有重排或扩增。未发现MSP基因家族周围各种限制性位点存在甲基化的证据,同样,也未观察到甲基化与表达之间的相关性。已克隆出该MSP基因家族的三个不同成员,并确定了它们的核苷酸序列。对成年雄性聚腺苷酸化mRNA构建的cDNA克隆文库进行差异筛选,得到45个雄性特异性克隆,其中32个是MSP基因的克隆。发现其中一个cDNA克隆包含MSP的完整核苷酸序列,包括部分5'前导序列和所有3'尾随序列。已分离出携带MSP基因拷贝的基因组克隆。该基因家族的至少一个成员是假基因。从基因组DNA克隆的MSP基因家族的另一个成员,除了一个5'侧翼序列外,还包含MSP的完整不间断结构序列,该侧翼序列含有一个具有经典TATA盒的启动子样区域、一个类似CAAT盒的序列和一个假定的核糖体结合序列。基因组和cDNA克隆的3'尾随序列包含一个AATAAA聚腺苷酸化位点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c622/368732/bf2f926e7ccf/molcellb00145-0150-a.jpg

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