Robinson M O, McCarrey J R, Simon M I
Division of Biology, California Institute of Technology, Pasadena 91125.
Proc Natl Acad Sci U S A. 1989 Nov;86(21):8437-41. doi: 10.1073/pnas.86.21.8437.
The gene encoding testis-specific phosphoglycerate kinase 2 (PGK; ATP:3-phospho-D-glycerate 1-phosphotransferase, EC 2.7.2.3) is expressed only in meiotic and haploid male germ cells. Transgenic mice containing an 8-kilobase human genomic PGK2 gene express the human gene in a tissue-specific and developmentally regulated manner. To determine the nature and location of sequences controlling this expression, transgenic mice with various lengths of the human PGK2 5' region fused to the chloramphenicol acetyltransferase (CAT) gene were analyzed for expression. A 323-base-pair region 5' to the coding region was found to contain information essential for both tissue-specific and developmentally regulated expression of the CAT reporter gene. Transgenic mice containing a PGK2/luciferase-coding construct were compared with mice containing an equivalent CAT construct. Luciferase gene expression was also testis-specific and was more sensitive than CAT gene expression, but otherwise regulation of the two reporter genes was similar in the germ cells of transgenic mice. Translation of both PGK2/CAT and PGK2/luciferase fusion genes was seen concurrently with the first detectable transcripts.
编码睾丸特异性磷酸甘油酸激酶2(PGK;ATP:3-磷酸-D-甘油酸1-磷酸转移酶,EC 2.7.2.3)的基因仅在减数分裂期和单倍体雄性生殖细胞中表达。含有8千碱基对人类基因组PGK2基因的转基因小鼠以组织特异性和发育调控的方式表达人类基因。为了确定控制这种表达的序列的性质和位置,分析了将不同长度的人类PGK2 5'区域与氯霉素乙酰转移酶(CAT)基因融合的转基因小鼠的表达情况。发现在编码区5'端的一个323碱基对区域包含CAT报告基因组织特异性和发育调控表达所必需的信息。将含有PGK2/荧光素酶编码构建体的转基因小鼠与含有等效CAT构建体的小鼠进行比较。荧光素酶基因表达也是睾丸特异性的,并且比CAT基因表达更敏感,但在转基因小鼠的生殖细胞中,这两种报告基因的调控在其他方面是相似的。PGK2/CAT和PGK2/荧光素酶融合基因的翻译与首次检测到的转录本同时出现。