版纳微型猪近交系白细胞介素6基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2015.08.002

›› 2015, Vol. 42 ›› Issue (8): 1925-1934.doi: 10.16431/j.cnki.1671-7236.2015.08.002

版纳微型猪近交系白细胞介素6基因克隆、生物信息学及组织表达分析

王配^1,2, 陈园园^1,2, 霍金龙^1,2, 霍海龙³, 王淑燕^1,2, 潘伟荣^1,2, 成文敏^1,2, 查星琴^1,2, 曾养志^1,2

1. 云南农业大学, 云南省版纳微型猪近交系重点实验室, 昆明 650201;
2. 云南农业大学动物科学技术学院, 昆明 650201;
3. 云南农业职业技术学院畜牧兽医系, 昆明 650031

收稿日期:2014-12-31 出版日期:2015-08-20 发布日期:2015-08-27
通讯作者: 霍金龙 E-mail:jinlonghuo973@163.com
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
基金资助:
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)

Cloning,Bioinformatics and Tissue Expression Analysis of IL-6 Gene in Banna Minipig Inbred Line

WANG Pei^1,2, CHEN Yuan-yuan^1,2, HUO Jin-long^1,2, HUO Hai-long³, WANG Shu-yan^1,2, PAN Wei-rong^1,2, CHENG Wen-min^1,2, ZHA Xing-qin^1,2, ZENG Yang-zhi^1,2

1. Key Laboratory of Banna Minipig Inbred Line of Yunnan Province, Yunnan Agricultural University, Kunming 650201, China;
2. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China;
3. Department of Husbandry and Veterinary, Yunnan Vocational and Technical College of Agriculture, Kunming 650031, China

Received:2014-12-31 Online:2015-08-20 Published:2015-08-27

摘要/Abstract

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23.88 ku,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。

关键词: 版纳微型猪近交系; 白细胞介素6; 免疫调节; 基因克隆; 组织表达

Abstract: Interleukin-6 (IL-6) is an important immune regulatory factor, and has great development 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 polymerase chain reaction (PCR) method, and had deposited the sequence into NCBI database and assigned to the accession No.JQ839263.Then we further studied this gene by bioinformatics analysis 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 domain and one transmembrance structure, 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 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 AF518322, 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 foundation for further insight into the expression and regulation of this gene of pig.

Key words: Banna minipig inbred line (BMI); IL-6; immune regulation; gene clone; tissue expression

中图分类号:

Q786

王配, 陈园园, 霍金龙, 霍海龙, 王淑燕, 潘伟荣, 成文敏, 查星琴, 曾养志. 版纳微型猪近交系白细胞介素6基因克隆、生物信息学及组织表达分析[J]. , 2015, 42(8): 1925-1934.

WANG Pei, CHEN Yuan-yuan, HUO Jin-long, HUO Hai-long, WANG Shu-yan, PAN Wei-rong, CHENG Wen-min, ZHA Xing-qin, ZENG Yang-zhi. Cloning,Bioinformatics and Tissue Expression Analysis of IL-6 Gene in Banna Minipig Inbred Line[J]. , 2015, 42(8): 1925-1934.

参考文献

[1] 李昌明,肖青,韩建魁.白细胞介素-6的临床应用进展[J].中国误诊学杂志,2007,7(1):30-32.
[2] Remington M C,Munson E L,Callister S M,et al.Interleukin-6 enhances production of anti-OspC immunoglobulin G2b borreliacidal antibody[J].Infect Immun,2001,69(7):4268-4275.
[3] Wu W C,Sun H W,Chen H T,et al.Circulating hematopoietic stem and progenitor cells are myeloid-biased in cancer patients[J].Proc Natl Acad Sci USA,2014,111(11):4221-4226.
[4] Chiarella S E,Soberanes S,Urich D,et al.β₂-adrenergic agonists augment air pollution-induced IL-6 release and thrombosis[J].J Clin Invest,2014,124(7):2935-2946.
[5] 庄晓明.肿瘤坏死因子-α、白细胞介素-6和白细胞介素-2受体与甲状腺肿瘤的相关性研究[J].中华实用诊断与治疗杂志,2010,24(1):32-33.
[6] Cies'lak M,Wojtczak A,Cies'lak M.Relationship between the induction of inflammatory processes and infectious diseases in patients with ischemic stroke[J].Acta Biochim Pol,2013,60(3):345-349.
[7] Wu M,Gao R,Meng M J,et al.Regulating effects of porcine interleukin-6 gene and CpG motifs on immune responses to porcine trivalent vaccines in mice[J].Res Vet Sci,2004,77(1):49-57.
[8] 许昌秦,李陕区.白细胞介素研究新进展[J].武警医学院学报,2007,16(2):191-194.
[9] Northemann W,Braciak T A,Hattori M,et al.Structure of the rat interleukin 6 gene and its expression in macrophage-derived cells[J].J Biol Chem,1989,264(27):16072-16082.
[10] Richards C D,Saklatvala J.Molecular cloning and sequence of porcine interleukin 6 cDNA and expression of mRNA in synovial fibriblasts in vitro[J].Cytokine,1991,3(4):269-276.
[11] Huo J L,Huo H L,Wang P,et al.The association of MC1R gene with coat color of Banna mini-pig inbred line (BMI)[J].J Anim Vet Adv,2012,11(4):503-508.
[12] 王配,霍金龙,李莲军.版纳微型猪近交系AQP3克隆、序列分析及组织表达[J].中国动物实验学报,2011,19(5):366-370.
[13] 李颖,霍金龙,潘伟荣,等.实时荧光定量RT-PCR法建立版纳微型猪近交系中厚皮创面转化生长因子表达体系[J].中国组织工程研究与临床康复,2011,15(7):1186-1190.
[14] 王淑燕,霍金龙,潘伟荣,等.版纳微型猪近交系GHR基因克隆及生物信息学分析[J].中国实验动物学报,2014,22(1):48-55.
[15] Qin C Y,Wang P,Wang S Y,et al.Complementary DNA cloning,sequence characterization and expression analyses of a novel gene MFN1 from Chinese Banna mini-pig inbred line (BMI)[J].Res J Biotechnol,2014,9(5):1-8.
[16] 张玲华,郭勇,田兴山,等.白细胞介素-6的分子生物学研究进展及其在畜禽业中的应用前景[J].中国畜牧兽医,2005,32(2):31-33.
[17] Kamimura D,Ishihara K,Hirano T.IL-6 signal transduction and its physiological roles:The signal orchestration model[J].Rev Physiol Biochem Pharmacol,2003,149:1-38.
[18] Scamurra R,Arriaga C,Sprunger L,et al.Regulation of interleukin-6 expression in porcine immune cells[J].J Interferon Cytokine Res,1996,16(4):289-296.
[19] 孟民杰,高荣,唐漫书,等.猪淋巴细胞白细胞介素-6基因的克隆和序列分析[J].四川大学学报,2002,39(3):557-560.
[20] Choi S E,Choi K M,Yoon I H,et al.IL-6 protects pancreatic islet beta cells from pro-inflammatory cytokines-induced cell death and functional impairment in vitro and in vivo[J].Transpl Immunol,2004,13(1):43-53.

版纳微型猪近交系白细胞介素6基因克隆、生物信息学及组织表达分析

Cloning,Bioinformatics and Tissue Expression Analysis of IL-6 Gene in Banna Minipig Inbred Line

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