Cloning and Bioinformatics Analysis of CYP1A1 of Cytochrome P450 Family from Banna Mini-pig Inbred Line
WANG Shuyan1,2, WANG Pei1,2, HUO Jinlong1,2, ZHA Xingqin1,2, PAN Weirong1,2, ZHANG Xia1,2, WANG Xuefei2
1. Key Laboratory of Banna Mini-pig Inbred Line of Yunnan Province, Yunnan Agricultural University, Kunming 650201, China; 2. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China
[Purpose] Our purpose was to clone sequence and analyze the character of the cytochrome P450 family 1 subfamily A member 1 gene (CYP1A1) from Banna mini-pig inbred line (BMI).[Methods] The complete CDS of CYP1A1 gene was cloned using RT-PCR method from BMI. Nucleotide and protein sequences of CYP1A1 were utilized to carry out bioinformatics analysis and construct the phylogenetic tree.[Results] The CDS length of BMI CYP1A1 is 1 551 bp in length which encodes a protein of 516 amino acids with molecular weight (Mw) 58.39 ku, and isoelectric point (pI) 7.83, and locates in the endoplasmic reticulum with 94.1% reliability. CYP1A1 protein contains a cytochrome P450 cysteine heme-iron ligand signature, an N-myristoylation site, a aidation site, two N-glycosylation sites, six protease C phosphorylation sites and five protein kinase Ⅱ phosphorylation sites. Phylogenetic analysis of 11 species showed that CYP1A1 of BMI (pig) had the closest relationship with these of cattle and goat.[Conclusion] The results will lay a foundation for further study on CYP1A1 gene function in BMI.
WANG Shuyan, WANG Pei, HUO Jinlong, ZHA Xingqin, PAN Weirong, ZHANG Xia, WANG Xuefei.Cloning and Bioinformatics Analysis of CYP1A1 of Cytochrome P450 Family from Banna Mini-pig Inbred Line[J] Journal of Yunnan Agricultural University, 2017,V32(3): 442-449
 NEBERT D W, ROE A L, DIETER M Z, et al. Role of the aromatic hydrocarbon receptor and[Ah] gene battery in the oxidative stress response, cell cycle control, and apoptosis[J]. Biochemical Pharmacology, 2000, 59(1):65.  GASPARI L, CHANG S S, SANTELLA R M, et al. Polycyclic aromatic hydrocarbon-DNA adducts in human sperm as a marker of DNA damage and infertility[J]. Mutation Research, 2003, 535(2):155.  MARINKOVIC' N, PASALIC' D, POTOCKI S. Polymorphisms of genes involved in polycyclic aromatic hydrocarbons' biotrans formation and atherosclerosis[J]. Biochemia Medica, 2013, 23(3):255.  ZHAN P, WANG Q, QIAN Q, et al. CYP1A1 MspI and exon7 gene polymorphisms and lung cancer risk:an updated meta-analysis and review[J]. Journal of Experimental & Clinical Cancer Research:CR, 2011, 30:99.DOI:10.1186/1756-9966-30-99.  陈文成,康熙雄,黄赞松,等.精子CYP1A1基因多态性与壮族人群少精症的关系[J].广东医学,2010,31(13):1669.DOI:10.3969/j.issn.1001-9448.2010.13.011.  杨家大,商海涛,魏泓.应用TaqMan定量方法研究巴马香猪CYP1A1、2A19、2E1 mRNA表达[J].中国实验动物学报,2011,19(6):534. DOI:10.3969/j.issn.1005-4847.2011.06.017.  KOJIMA M, SEKIMOTO M, DEGAWA M. Androgen-mediated down-regulation of CYP1A subfamily genes in the pig liver[J]. Journal of Endocrinology, 2010, 207(2):203. DOI:10.1677/JOE-10-0160.  徐杰,赵为民,任守文,等.CYP1A1与PPAR-γ在猪肺炎支原体感染炎性反应调控中的作用关系[J].畜牧兽医学报,2016,47(3):574. DOI:10.11843/j.issn.0366-6964.2016.03.021.  HUO J L, WANG P, ZHAO Y E, et al. Molecular cloning, mRNA expression and characterization of a novel FAIM1 gene from Chinese banna mini-pig inbred line (BMI)[J]. Journal of Animal and Veterinary Advances, 2012, 11(8):1080.  WANG P, HUO H L, WANG S Y, et al. Cloning, sequence characterization, and expression patterns of members of the porcine TSSK family[J]. Genetics and Molecular Research:GMR, 2015, 14(4):14908.DOI:10.4238/2015.October.18.56.  DEGTYARENKO K N, ARCHAKOV A I. Molecular evolution of P450 superfamily and P450-containing monooxygenase systems[J]. FEBS Letters, 1993, 332(1/2):1.  MOREL Y, MERMOD N, BAROUKI R. An autoregulatory loop controlling CYP1A1 gene expression:role of H2O2 and NFI[J]. Molecular and Cellular Biology, 1999, 19(10):6825.  SCHLEZINGER J J, WHITE R D, STEGEMAN J J. Oxidative inactivation of cytochrome P-4501A (CYP1A) stimulated by 3,3',4,4'-tetrachlorobiphenyl:production of reactive oxygen by vertebrate CYP1As[J]. Molecular Pharmacology, 1999, 56(3):588.  MOLCAN T, SWIGONSKA S, ORLOWSKA K, et al. Structural-functional adaptations of porcine CYP1A1 to metabolize polychlorinated dibenzo-p-dioxins[J]. Chemosphere, 2017, 168:205.DOI:10.1016/j.chemosphere.2016.10.035.  THEILE D, ALLENDORF D, KÖHLER B C, et al. Obatoclax as a perpetrator in drug-drug interactions and its efficacy in multidrug resistance cell lines[J]. The Journal of Pharmacy and Pharmacology, 2015, 67(11):1575.DOI:10.1111/jphp.12455.  FANG X M, ZHAO W M, XU J, et al. CYP1A1 mediates the suppression of major inflammatory cytokines in pulmonary alveolar macrophage (PAM) cell lines caused by Mycoplasma hyponeumoniae[J]. Developmental and Comparative Immunology, 2016, 65:132.DOI:10.1016/j.dci.2016.06.023.  GUENGERICH F P. Reactions and significance of cytochrome P-450 enzymes[J]. The Journal of Biological Chemistry, 1991, 266(16):10019.  孙佳,陆苑,潘洁,等.参芎葡萄糖注射液对肝脏6种亚型微粒体细胞色素P450酶的抑制作用[J].贵州医科大学学报,2016,41(7):775. DOI:10.19367/j.cnki.1000-2707.2016.07.007.