Elizur A, Zmora N, Rosenfeld H, Meiri I, Hassin S, Gordin H, Zohar Y
Israel Oceanographic and Limnological Research, National Center for Mariculture, Eilat, Israel.
Gen Comp Endocrinol. 1996 Apr;102(1):39-46. doi: 10.1006/gcen.1996.0044.
Full-length cDNA sequences encoding the beta-subunits of the gonadotrophins GtHI and GtHII were isolated, cloned, and sequenced from a single gilthead seabream (Sparus aurata) pituitary using RACE PCR. Beta-GtHI and beta-GtHII degenerate PCR primers were designed according to regions of high amino acid sequence homology between the chum salmon and the bonito beta-GtHI or beta-GtHII. DNA sequence analysis of the cloned PCR products confirmed the presence of the predicted complete coding region as well as 5' and 3' untranslated sequences. The deduced amino acid sequences of beta-GtHI and beta-GtHII from the gilthead seabream were compared to GtH sequences from a number of teleosts. Seabream beta-GtHI shows the highest homology to the bonito beta-GtHI (76%) and the striped bass (74%), while seabream beta-GtHII is 98% homologous to the yellow fin porgy, 87% homologous to the bonito, and 93% homologous to the striped bass beta-GtHII. Northern blot analysis showed the transcripts of the gilthead seabream beta-GtHI and beta-GtHII to be about 700 and 740 bases, respectively. During the spawning season, beta-GtHI is expressed at higher levels in males than in females while beta-GtHII is expressed at similar levels in both sexes.
利用RACE PCR技术，从一条金头鲷（Sparus aurata）垂体中分离、克隆并测序了编码促性腺激素GtHI和GtHIIβ亚基的全长cDNA序列。根据马苏大麻哈鱼与鲣鱼的β-GtHI或β-GtHII之间氨基酸序列高度同源的区域，设计了β-GtHI和β-GtHII简并PCR引物。对克隆的PCR产物进行DNA序列分析，证实了预测的完整编码区以及5'和3'非翻译序列的存在。将金头鲷β-GtHI和β-GtHII推导的氨基酸序列与多种硬骨鱼的GtH序列进行了比较。金头鲷β-GtHI与鲣鱼β-GtHI（76%）和条纹鲈（74%）的同源性最高，而金头鲷β-GtHII与黄鳍鲷的同源性为98%，与鲣鱼的同源性为87%，与条纹鲈β-GtHII的同源性为93%。Northern印迹分析表明，金头鲷β-GtHI和β-GtHII的转录本分别约为700和740个碱基。在产卵季节，β-GtHI在雄性中的表达水平高于雌性，而β-GtHII在两性中的表达水平相似。
