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首页> 外文期刊>Biochimica et biophysica acta: international journal of biochemistry and biophysics >Assignment of chromosomal locus and evidence for alternatively spliced mRNAs of a human sperm membrane protein (hSMP-1).
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Assignment of chromosomal locus and evidence for alternatively spliced mRNAs of a human sperm membrane protein (hSMP-1).

机译：染色体基因座的分配以及人类精子膜蛋白（hSMP-1）选择性剪接的mRNA的证据。

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摘要

The gene (HSD-1) coding a human sperm membrane protein (hSMP-1) was isolated from a human testis cDNA expression library using antibodies found in the serum of an infertile woman. HSD-1 was localized to a single locus on chromosome 9 and assigned to band 9p12-p13 by fluorescent in situ hybridization (FISH) mapping and DAPI (4,6-diamidino-2-phenylindole) banding, using rat/human somatic cell hybrids and metaphase chromosomes of human lymphocytes. In rescreening a testis lambdagt10 cDNA expression library, the full-length cDNA (HSD-1) and several truncated cDNAs with heterologous regions were isolated from positive clones. The heterology consisted of deletion, insertion and alteration of the 5'-end. These heterologous truncated fragments may be produced by alternative splicing of mRNAs. Two recombinant prokaryotic expression vectors were constructed with one of the heterologous fragment (clone #26) with and without the alternative 5'-end. Escherichia coli transfected with the construct containing the alternative 5'-end failed to produce the recombinant product, whereas those transfected with the vector lacking the 5'-end produced hSMP-1. DNASIS analysis of the structure of #26 mRNA suggests that the 5'-end has a stable secondary configuration that may maintain the mRNA in an inactivated state, whereby hindering its translation and preventing the expression of the gene.
机译：使用在不育妇女血清中发现的抗体，从人睾丸cDNA表达文库中分离出编码人精子膜蛋白（hSMP-1）的基因（HSD-1）。使用大鼠/人类体细胞杂种，将HSD-1定位于9号染色体上的单个基因座，并通过荧光原位杂交（FISH）定位和DAPI（4,6-diamidino-2-phenylindole）带将其分配给9p12-p13带。和人类淋巴细胞的中期染色体。在重新筛选lambdagt10睾丸cDNA表达文库时，从阳性克隆中分离出全长cDNA（HSD-1）和具有异源区域的多个截短cDNA。异质性包括5'末端的缺失，插入和改变。这些异源截短的片段可以通过mRNA的可变剪接产生。用具有和不具有替代5'端的异源片段之一（克隆＃26）构建两个重组原核表达载体。用含有替代的5'-末端的构建体转染的大肠杆菌不能产生重组产物，而用缺少5'-末端的载体转染的大肠杆菌产生了hSMP-1。 DNASIS分析＃26 mRNA的结构表明5'端具有稳定的二级构型，可以将mRNA保持在灭活状态，从而阻碍其翻译并阻止该基因的表达。

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