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作 者:杜玉杰[1] 侯怡铃[2] 田丽[3] 侯万儒[2]
机构地区:[1]湛江师范学院基础教育学院生化系,广东湛江524048 [2]西华师范大学生命科学院,四川南充637002 [3]湛江师范学院生命科学与技术学院,广东湛江524048
出 处:《西南大学学报(自然科学版)》2012年第6期50-56,共7页Journal of Southwest University(Natural Science Edition)
基 金:国家自然科学基金资助项目(30570275);四川省基础应用基金资助项目(2009JY0061)
摘 要:该研究运用RT-PCR技术,首次从大熊猫Ailuropoda melanoleuca的肌肉组织总RNA中成功克隆了AP2S1基因的编码序列,并对其进行了全面分析.结果表明:大熊猫AP2S1基因编码序列克隆片段全长为512bp,开放阅读框(ORF)为429bp,编码142个氨基酸的蛋白质,该蛋白的相对分子质量为1.7×104,等电点pI为5.82,含有4种类型共5个功能位点,即1个蛋白激酶C磷酸化位点、2个酪蛋白激酶Ⅱ磷酸化位点、1个N-酰基化位点和1个网格蛋白调节素小肽链标签;且该蛋白在哺乳动物网状细胞中的半寿期(half-life)大于30h,不稳定指数(in-stability index)为26.99,属于稳定蛋白.进一步分析发现,大熊猫AP2S1基因与已报道的人、苏门答腊猩猩、牛、褐家鼠和小家鼠5个哺乳动物物种的编码序列及其编码的氨基酸序列具有很高的相似性,表明该基因及其编码蛋白在物种漫长的进化过程中极其保守.The cDNA of AP2S1 was cloned successfully for the first time from the muscle tissue of giant panda (Ailuropoda rnelanoleuca) using RT-PCR technology and was subsequently sequenced and prelimi narily analyzed. The cDNA fragment cloned was shown to be 512 bp in size, containing an open reading frame of 429 bp. The deduced protein sequence showed that the protein was composed of 142 amino acids and its estimated molecular weight was 17 017.7 with a pI of 5.82. Topology prediction showed that it had one protein kinase C phosphorylation site, two casein kinase II phosphorylation sites, one N-myristoyla- tion site, one amidation site and one clathrin adaptor complexes small chain signature in the AP2S1 protein of A. melanoleuca. The protein was identified as a stable protein, whose estimated half-life in mammalian reticulocytes was 30 hours, with an instability index of 26.99. Alignment analysis indicated that the nucleotide sequence and the deduced amino acid sequence of A. melanoleuca shared high similarity to those of some mammals, including Homo sapiens, Pongo abeli , Bos taurus, Rattus norvegicus and Mus musculus , thus suggesting that this gene of A. melanoleuca and the protein it encodes are highly conserved during the evolutionary process.
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