Otani T, Otani F, Krych M, Chaplin D D, Boime I
Department of Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63110.
J Biol Chem. 1988 May 25;263(15):7322-9.
To identify the promoter sequence(s) of the CG beta gene, genomic fragments derived from a cosmid containing the CG beta gene family were transfected into mouse Y1 adrenal cortical cells. Using this system, we showed that the CG beta genes 5, 3, and 8 have functional promoters, the basal element of which in the case of CG beta 5, was within 78 base pairs 5' ward from the CAP site. The size of the CG beta transcripts and identity of the transcription start site was the same for CG beta mRNA synthesized in Y1 cells as in first trimester placenta. The promoter region identified by this system was also capable of driving the chloramphenicol acetyltransferase gene when transfected stably into choriocarcinoma cells. Chloramphenicol acetyltransferase constructs bearing variable length of 5'-flanking sequences from CG beta 5 and transfected into trophoblast cells suggest the presence of regulatory sequences within 700 base pairs from the CAP site. The information obtained here provide a foundation for studies of analyzing trans-acting placental and pituitary proteins to the defined CG beta promoter region.
为了鉴定绒毛膜促性腺激素β(CGβ)基因的启动子序列,将来自含有CGβ基因家族的黏粒的基因组片段转染到小鼠Y1肾上腺皮质细胞中。利用该系统,我们发现CGβ基因5、3和8具有功能性启动子,就CGβ5而言,其基本元件位于距帽位点5'端78个碱基对内。Y1细胞中合成的CGβmRNA的转录本大小和转录起始位点与孕早期胎盘相同。该系统鉴定出的启动子区域在稳定转染到绒毛膜癌细胞中时也能够驱动氯霉素乙酰转移酶基因。携带来自CGβ5的可变长度5'侧翼序列并转染到滋养层细胞中的氯霉素乙酰转移酶构建体表明在距帽位点700个碱基对内存在调控序列。此处获得的信息为分析与确定的CGβ启动子区域相互作用的胎盘和垂体反式作用蛋白的研究奠定了基础。