The multisubunit NADH ubiquinone oxidoreductase (complex I) is the first enzyme complex in the electron transport chain of mitochondria. The iron-sulfur protein (IP) fraction is made up of 7 subunits, including NDUFS6 (NADH-ubiquinone oxidoreductase Fe-S protein 6). In order to understand the giant panda’s NDUFS6 gene and coding of protein using RT-PCR combined with in silico cloning, we isolated and sequenced the cDNA encoding NADH-ubiquinone oxidoreductase iron-sulfur protein 6 of the giant panda (Ailuropoda melanoleuca).The deduced protein sequence showed that the protein is composed of 124 amino acids and the estimated molecular weight of the NDUFS6 protein is 13.632 kDa with an isoelectric point (PI) of 9.51. Alignment analysis reveals that the deduced protein sequence shares 85.6 %,82.4 %,76.6 % and 76.1 % homology with that of Homo sapiens, Bos taurus, Rattus norvegicus and Mus musculus, respectively. In particular, there are 2 Protein kinase C phosphorylation sites (14SGK16 and51SRK53) in the NDUFS6 protein of the giant panda.
Scleroderma is a connective tissue disease characterized by the fibrosisization of skin and internal organs.In this study,the genomic sequence and cDNA of Sjogren’s syndrome/scleroderma autoantigen 1(SSSCA1)gene of the giant panda was cloned successfully using RT-PCR.The cDNA fragment cloned was 642 bp in length,and contained an open reading frame (ORF) of 600 bp encoding 199 amino acids.The genomic sequence was 1 262 bp,containing four exons and three introns. The deduced amino acid sequence showed that this protein was composed of 199 amino acids and its estimated molecular weight was 21.53841 kDa with a pI of 5.14. Three different patterns of functional sites were found:one protein kinase C phosphorylation site,five casein kinase Ⅱ phosphorylation sites,and three N-myristoylation sites. The SSSCA1 gene of the giant panda were highly homologous to those of some other mammals.
本研究运用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RPS24(Ribosomal Protein S24)基因的表达序列,并对其进行了测序及初步分析.结果表明:大熊猫RPS24基因的表达序列全长为431bp,开放阅读框(ORF)为399bp,编码132个氨基酸的蛋白质,该蛋白的分子量为15.3251kD,pI为10.92,含有7种类型共14个功能位点:即1个N-糖基化位点、2个cAMP和cGMP依赖性蛋白激酶磷酸化位点、6个蛋白激酶C磷酸化位点、1个酪蛋白激酶Ⅱ磷酸化位点、3个N-酰基化位点、1个酰胺化位点及1个RPS24e sig-nature.进一步分析发现,大熊猫RPS24基因与已报道的人、牛、苏门答腊猩猩、褐家鼠和小家鼠5个哺乳动物物种的表达序列及其编码的氨基酸序列具有很高的相似性:编码序列同源性分别为94.1%、92.4%、94.7%、89.9%和90.4%;与人RPS24蛋白的氨基酸序列同源性为98.48%,其余均为99.24%.
