Ho K C, Quarmby V E, French F S, Wilson E M
Laboratories for Reproductive Biology, University of North Carolina, Chapel Hill 27599.
J Biol Chem. 1992 Jun 25;267(18):12660-7.
Complementary DNA (cDNA) that codes for a major androgen-dependent secretory protein of rat coagulating gland and dorsal prostate, dorsal protein 1 (DP1), was isolated by molecular cloning. Recombinant DP1 cDNA clones were identified from a bacteriophage lambda gt11 rat coagulating gland expression library using an affinity purified polyclonal antibody. Amino acid sequence deduced from DNA contained sequences identical with several DP1 cyanogen bromide cleavage fragments. Northern blot hybridization of poly(A) RNA isolated from intact rat dorsal prostate and coagulating gland revealed a predominant messenger RNA (mRNA) species of approximately 3200 nucleotides. Tissue-specific expression of DP1 mRNA was indicated by the absence of DP1 mRNA in ventral prostate and other tissues of the rat. Expression of DP1 mRNA was androgen-dependent, decreasing approximately 80% 7 days after castration and increasing rapidly following androgen replacement. Southern blot analysis of restriction enzyme-digested rat DNA indicated that DP1 is encoded by a single gene and that no major genomic rearrangements accounted for its lack of expression in the dorsal prostate-derived rat Dunning tumor. Sequence comparisons revealed that rat prostate DP1 shares sequence identity with Factor XIIIa and tissue transglutaminase, including the active center, GQCWVF, indicating that DP1 is a member of the transglutaminase gene family.
通过分子克隆分离出了编码大鼠凝固腺和背侧前列腺主要雄激素依赖性分泌蛋白背侧蛋白1(DP1)的互补DNA(cDNA)。使用亲和纯化的多克隆抗体从噬菌体λgt11大鼠凝固腺表达文库中鉴定出重组DP1 cDNA克隆。从DNA推导的氨基酸序列包含与几个DP1溴化氰裂解片段相同的序列。从完整大鼠背侧前列腺和凝固腺分离的聚腺苷酸RNA(poly(A) RNA)的Northern印迹杂交显示,主要的信使RNA(mRNA)种类约为3200个核苷酸。大鼠腹侧前列腺和其他组织中不存在DP1 mRNA,表明DP1 mRNA具有组织特异性表达。DP1 mRNA的表达是雄激素依赖性的,去势7天后降低约80%,雄激素替代后迅速增加。对限制性内切酶消化的大鼠DNA进行Southern印迹分析表明,DP1由单个基因编码,并且在源自背侧前列腺的大鼠邓宁肿瘤中其缺乏表达并非由主要的基因组重排所致。序列比较显示,大鼠前列腺DP1与因子XIIIa和组织转谷氨酰胺酶具有序列同一性,包括活性中心GQCWVF,表明DP1是转谷氨酰胺酶基因家族的成员。