Saunders M C, Deakin J, Harrison G A, Curlewis J D
Department of Physilogy and Pharmacology, University of Queensland, Australia.
Gen Comp Endocrinol. 1998 Jul;111(1):68-75. doi: 10.1006/gcen.1998.7090.
The full-length nucleotide sequence for the cDNA of growth hormone (GH) from the marsupial brushtail possum (Trichosurus vulpecula) was determined using reverse transcription followed by polymerase chain reaction (RT-PCR) and the 5'/3' rapid amplification of cDNA ends (RACE) technique. Sequence information showed that the possum GH cDNA was 831 base pairs (bp) in length, including the 5'untranslated region (60 bp), the signal peptide (75 bp), the mature protein (573 bp, including stop codon), and the 3' untranslated region (123 bp). At both the nucleotide and the amino acid level (deduced from nucleotide sequence), there was a high degree of sequence identity with pig and horse. These species were similar at 82.8 and 83.0% of bases at the nucleotide level and 91.6 and 91.1% at the amino acid level, respectively. Northern analysis showed that GH mRNA is present in the pituitary gland and was similar in size to that seen in other mammals (approximately equal to 0.9 kb). Analysis of molecular evolution of GH in the possum indicated that the rate of evolution is relatively slow (0.4 substitutions/ amino acid site/year x 10(9) and typical of that seen for nonprimate mammals, which exhibit rates ranging between 0.2 and 1.3 substitutions/amino acid site/year x 10(9).
采用逆转录聚合酶链反应(RT-PCR)及5'/3' cDNA末端快速扩增(RACE)技术,测定了有袋动物帚尾袋貂(Trichosurus vulpecula)生长激素(GH)cDNA的全长核苷酸序列。序列信息显示,袋貂GH cDNA长度为831个碱基对(bp),包括5'非翻译区(60 bp)、信号肽(75 bp)、成熟蛋白(573 bp,包括终止密码子)和3'非翻译区(123 bp)。在核苷酸和氨基酸水平(从核苷酸序列推导)上,与猪和马均有高度的序列同一性。这些物种在核苷酸水平上分别有82.8%和83.0%的碱基相似,在氨基酸水平上分别为91.6%和91.1%。Northern分析表明,GH mRNA存在于垂体中,其大小与其他哺乳动物所见相似(约0.9 kb)。对袋貂GH分子进化的分析表明,其进化速率相对较慢(0.4个替换/氨基酸位点/年×10^9),这是典型的非灵长类哺乳动物的进化速率,其范围在0.2至1.3个替换/氨基酸位点/年×10^9之间。