Takahashi N, Kikuyama S, Gen K, Maruyama O, Kato Y
Department of Biology, School of Education, Waseda University, Tokyo, Japan.
J Mol Endocrinol. 1992 Dec;9(3):283-9. doi: 10.1677/jme.0.0090283.
A GH cDNA was specifically amplified from cDNAs constructed from total RNA of bullfrog (Rana catesbeiana) adenohypophyses employing the DNA polymerase chain reaction. Sequencing analysis revealed that the cDNA clone thus obtained was 654 bp in length, and included an open reading frame encoding the entire sequence of mature GH, with its signal peptide. Slight discrepancies were noted between the deduced amino acid sequence and that determined by direct protein sequencing of purified bullfrog GH or that deduced from the nucleotide sequence reported previously. The length of the bullfrog GH mRNA was estimated to be about 1.2 kb by Northern blot analysis. Homologies of nucleotide and amino acid sequences between GH and prolactin of bullfrog origin were 48% and 26% respectively. Using the cDNA as a probe, the content of GH mRNA in the pituitary of larval and adult bullfrogs was measured. GH mRNA levels were relatively low at the preclimax stage, and rose markedly during climax. In juvenile frogs, GH mRNA levels in the pituitary were extremely high and declined towards adulthood. This finding suggests that the increase in plasma and pituitary GH levels reported previously accompanies the increase in GH synthesis.
利用DNA聚合酶链式反应,从牛蛙(Rana catesbeiana)腺垂体总RNA构建的cDNA中特异性扩增出生长激素(GH)cDNA。测序分析表明,所获得的cDNA克隆长度为654 bp,包含一个编码成熟GH完整序列及其信号肽的开放阅读框。推导的氨基酸序列与通过纯化的牛蛙GH直接蛋白质测序确定的序列或先前报道的核苷酸序列推导的序列之间存在轻微差异。通过Northern印迹分析估计牛蛙GH mRNA的长度约为1.2 kb。牛蛙来源的GH与催乳素之间的核苷酸和氨基酸序列同源性分别为48%和26%。以该cDNA为探针,测定了幼体和成体牛蛙垂体中GH mRNA的含量。在高潮前期,GH mRNA水平相对较低,在高潮期显著升高。在幼蛙中,垂体中的GH mRNA水平极高,并在成年期下降。这一发现表明,先前报道的血浆和垂体GH水平的升高伴随着GH合成的增加。