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基因和相应的蛋白序列进行了生物信息学分析,结果显示该基因编码212个氨基酸,分子质量为23.88ku,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 IL-6与GenBank登录号为NM_214399和DQ832259的核苷酸序列完全相同,与GenBank登录号为AF309651和AF518322的核苷酸序列各存在1处碱基差异;多物种氨基酸序列比对及系统进化分析结果表明,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,结果发现在肺脏、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。
Interleukin-6 （IL-6） is an important immune regulatory factor,and has great develop- ment prospects in the application of adjuvants. Using the Banna minipig inbred line （BMI） as the experimental animal,we successfully cloned the coding region of pig IL-6 gene through polymer- ase chain reaction （PCR） method, and had deposited the sequence into NCBI database and as- signed to the accession No. JQ839263. Then we further studied this gene by bioinformaties analy- sis method. The results showed that BMI IL-6 encoded 212 amino acids with a predicted molecular weight of 23.88 ku,the isoelectric point （pI） was 6.66 and a signal peptide in N-terminal. It was a nuclear protein with 89 ~ probability. In addition, BMI IL-6 contained one conserved structure do- main and one transmemhrance structure, and its N-terminal and C-terminal were both hydropho- hie. Comparing the sequence of the coding region of BMI IL-6 gene with the download 4 IL-6 gene sequences of pig,the result showed BMI was identical with GenBank accession No. NM_214399 and DQ832259, only had one base pair difference with GenBank accession No. AF309651 and AFS18322,respectively. Multi-species amino acid sequence alignment and phylogenetic analysis demonstrated that BMI IL-6 showed the closest relationship with camel. Finally,we analyzed IL-6 gene mRNA expression in 12 important tissues of BMI through PCR method,the results showed that BMI IL-6 gene was highly expressed in lung, skin and muscle of BMI. These data laid a foun- dation for further insight into the expression and regulation of this gene of pig.
出处
《中国畜牧兽医》
CAS
北大核心
2015年第8期1925-1934,共10页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31160439、31460580)
云南省教育厅科学研究基金项目(2011C191)
作者简介
王配（1987-），女，河南上蔡人，博士生，实验师，研究方向：动物分子遗传学，E—mail：peipei99999@126．com
陈园园（1990-），男。山东东平人，硕士生，研究方向：动物分子遗传学，E-mail：chenyuanyuan9030@163．com
通信作者：霍金龙（1975-），男，山西五寨人，博士，高级实验师，硕士生导师，主要从事动物分子生物学方面的研究，E-mail：jinlonghu0973@163．com
