McDonald C, Williams L, McTurk P, Fuller F, McIntosh E, Higgins S
Nucleic Acids Res. 1983 Feb 25;11(4):917-30. doi: 10.1093/nar/11.4.917.
Under the influence of testosterone, rat seminal vesicles synthesise large amounts of a tissue specific protein, S. Recombinant lambda clones have been isolated containing overlapping sequences covering a 27.5 kilo base region of the rat genome within which the gene for protein S is located. Recombinant plasmids bearing cDNA sequences for protein S were constructed in pBR328. One (pcS2) contains a 690 nucleotide insert and is probably full length. Detailed restriction maps of the S-gene are presented and the structure was confirmed by analysis of R-loops and heteroduplexes. The S-gene covers a 2 kbp region of the genome and consists of a 5' intron (490 bp) separating a leading exon (120 bp) containing the 5' untranslated region from a central exon (310 bp) containing most of the coding sequence and part of the 3' untranslated region. A larger intron (1100 bp) lies within the 3' untranslated region. The cloned gene is representative of the native gene but the S gene may be heterogeneous. Using pcS2, the hormonal control of S-specific mRNA was examined and a pronounced differential response to testosterone was observed.
在睾酮的影响下,大鼠精囊合成大量组织特异性蛋白S。已分离出重组λ克隆,其包含覆盖大鼠基因组27.5千碱基区域的重叠序列,蛋白S的基因位于该区域内。在pBR328中构建了携带蛋白S cDNA序列的重组质粒。其中一个(pcS2)含有一个690个核苷酸的插入片段,可能是全长的。给出了S基因的详细限制性图谱,并通过R环和异源双链体分析证实了其结构。S基因覆盖基因组的2千碱基区域,由一个5'内含子(490 bp)组成,该内含子将包含5'非翻译区的前导外显子(120 bp)与包含大部分编码序列和部分3'非翻译区的中央外显子(310 bp)分开。一个较大的内含子(1100 bp)位于3'非翻译区内。克隆的基因代表天然基因,但S基因可能是异质的。使用pcS2,研究了S特异性mRNA的激素调控,并观察到对睾酮有明显的差异反应。