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huojinlong8610金虫 (小有名气)
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[求助]
润色英文摘要
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摘 要：白细胞介素6（IL-6）是机体重要的免疫调节因子，在佐剂的应用方面具有很好的发展前景。本研究以版纳微型猪近交系（BMI）为实验动物，通过RT-PCR法获得IL-6基因编码区序列，提交GenBank数据库，登录号为“JQ839263”。对BMI IL-6基因和相应的蛋白质序列进行了多重生物信息学分析，结果显示该基因编码212个氨基酸，蛋白分子量（Mw）为23.88 kD，等电点（pI）为6.66，存在一个保守结构域，一个跨膜结构，N端和C端均疏水，且在N端存在信号肽，有89%的可能性定位于细胞核。将BMI的IL-6基因编码区序列与NCBI下载到的4条猪IL-6基因序列进行比对结果显示BMI IL6与NM_214399和DQ832259核酸序列完全相同，与AF309651和AF518322各存在1处碱基差异；多物种氨基酸序列比对及系统进化分析表明，BMI与骆驼亲缘关系最近。同时利用半定量PCR法确定了IL-6基因mRNA在BMI 12个组织中的表达量，发现在肺、皮肤、肌肉中表达量较高。本研究为进一步研究猪IL-6基因的表达调控奠定了理论基础。 Abstract：The interleukin-6 (IL-6) is an important immune regulatory factor, and has great development prospects in the application of adjuvants. Using the Banna Mini-pig Inbred Line (BMI) as the experimental animal, we successfully cloned the coding region of porcine IL-6 gene through the reverse transcriptase polymerase chain reaction (RT-PCR) method, and had deposited the sequence into NCBI database and assigned to the accession No.: JQ839263. Then we further study this gene by multiple bioinformatics analyses method. Results showed that BMI IL-6 encoded 212 amino acids with a predicted molecular weight of 23.88 kD and the isoelectric point (pI) 6.66 and a signal peptide in N-terminal. It is a nuclear protein with a 89% probability. In addition, BMI IL-6 contained one conserved domain and its N-terminal and C-terminal were both hydrophobic. Comparing the sequence of the coding region of BMI IL-6 gene with the download 4 pig IL-6 gene sequences, the result shows BMI is identical with NM_214399 and DQ832259, only had 1 base pair difference with AF309651 and AF518322 respectively. Multi-species amino acid sequence alignment and phylogenetic analysis showed that BMI has a closest relationship with camel. Finally, we analysed IL-6 gene mRNA expression in 12 important tissues of BMI through RT-PCR method, results showed that BMI IL-6 gene was highly expressed in lung, skin and muscle of BMI adult pig. These data serve as a foundation for further insight into the expression and regulation of this porcine gene. |
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