香猪卵巢StAR和CYP11A1基因的差异表达研究

doi:10.16431/j.cnki.1671-7236.2018.05.002

《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (5): 1137-1144.doi: 10.16431/j.cnki.1671-7236.2018.05.002

香猪卵巢StAR和CYP11A1基因的差异表达研究

毛宁^1,2, 王嘉福^2,3, 张福平⁴, 唐靓婷¹, 阮亦麒⁴, 易凡利⁴, 冉雪琴⁴

1. 贵州大学生命科学学院, 贵阳 550025;
2. 铜仁学院, 铜仁 554300;
3. 贵州大学农业生物工程研究院, 贵阳 550025;
4. 贵州大学动物科学学院, 贵阳 550025

收稿日期:2017-11-10 出版日期:2018-05-20 发布日期:2018-05-19
通讯作者: 王嘉福, 冉雪琴 E-mail:jfwang@gzu.edu.cn;rqran@gzu.edu.cn
作者简介:毛宁(1990-),男,贵州惠水人,硕士生,研究方向:动物分子生物学,E-mail:1364934005@qq.com
基金资助:
国家高技术研究发展计划（863计划）课题（2013AA102503）；国家自然科学基金（31672390）；贵州省农业攻关项目（黔科合支持[2017]2585、[2017]2587）；贵州省“百”层次创新型人才项目（黔科合人才2016-4012号）

Study on Differential Expression of StAR and CYP11A1 Genes in Xiang Pig Ovary

MAO Ning^1,2, WANG Jiafu^2,3, ZHANG Fuping⁴, TANG Liangting¹, RUAN Yiqi⁴, YI Fanli⁴, RAN Xueqin⁴

1. College of Life Sciences, Guizhou University, Guiyang 550025, China;
2. Tongren University, Tongren 554300, China;
3. Institute of Agricultural Bioengineering, Guizhou University, Guiyang 550025, China;
4. College of Animal Science, Guizhou University, Guiyang 550025, China

Received:2017-11-10 Online:2018-05-20 Published:2018-05-19

摘要/Abstract

摘要：

为了揭示香猪繁殖调控的分子机制，本试验结合生物信息学分析方法对StAR和CYP11A1基因在香猪卵巢组织中的差异表达进行分析。运用实时荧光定量PCR方法检测高、低产群体6月龄香猪卵巢组织中StAR和CYP11A1基因的表达量变化；从香猪基因组中克隆两个基因的启动子区域，分析两个基因表达差异的分子机制。高通量转录组测序和实时荧光定量PCR检测结果均显示，高产香猪两个基因的表达量较高，启动子区序列长分别为1 144与970 bp。StAR基因启动子区结合转录因子ER、COUP、Sp1等34种72个，转录起始位点上游68~698 bp存在8个变异位点，但不影响转录因子的结合；与CYP11A1基因启动子区相结合的转录因子为Sp1、PU.1、C/EBPdel等30种75个，转录起始位点上游12~593 bp存在5个变异位点，其中69 bp处检测到C-T变异，高产香猪的T等位基因检出频率为17%，低产香猪为33%；对预测结果整合分析，T等位基因丢失了转录因子Sp1的结合位点。高、低产香猪之间StAR和CYP11A1基因的表达存在差异，CYP11A1基因启动子区69 bp处的变异可能是基因表达量差异的原因之一。

关键词: 香猪; 卵巢; 实时荧光定量PCR; StAR基因; CYP11A1基因; 启动子

Abstract:

In order to reveal the molecular mechanism of Xiang pig reproduction, StAR and CYP11A1 genes of ovary in Xiang pigs were analyzed.Chosing two groups of Xiang pig with high and low litter sizes as samples,the expression level of two genes were further detected from ovary by Real-time PCR method.And the promoter regions of two genes were cloned and sequenced from the genomic DNA.The results showed that both of FPKM values from the RNA-seq for transcriptomes and the mRNA levels by Real-time PCR in the group with high litter size were much higher than the other one.In the promoter region of StAR gene with 1 144 bp,the binding elements of a total of 72 transcription factors (TF,34 kinds) were deduced,including ER,COUP and Sp1,etc.About 8 SNP sites were determined from sequence of 68 to 698 bp upstream of the transcription initiation site.All of SNP sites were outside of the binding sites for TFs.For the promoter of CYP11A1 gene in 970 bp,there presented 75 TFs (30 kinds),such as Sp1,PU.1 and C/EBPdel.About 5 SNP sites were detected in the region of 12 to 593 bp upstream of the start site,in which a polymorphism site of C-T at 69 bp would change the binding of SP1.The frequency of T allele was 17% and 33% in Xiang pigs with high or low litter sizes,respectively.The SNP site at 69 bp in the promoter region might be a reason for the different expression of CYP11A1 gene in pig ovary.

Key words: Xiang pig; ovary; Real-time PCR; StAR gene; CYP11A1 gene; promoter

中图分类号:

S828

毛宁, 王嘉福, 张福平, 唐靓婷, 阮亦麒, 易凡利, 冉雪琴. 香猪卵巢StAR和CYP11A1基因的差异表达研究[J]. 《中国畜牧兽医》, 2018, 45(5): 1137-1144.

MAO Ning, WANG Jiafu, ZHANG Fuping, TANG Liangting, RUAN Yiqi, YI Fanli, RAN Xueqin. Study on Differential Expression of StAR and CYP11A1 Genes in Xiang Pig Ovary[J]. , 2018, 45(5): 1137-1144.

参考文献

[1] 张笑,苏艳,杨世彬,等.从江香猪抑制素α基因克隆及其卵巢表达研究[J].中国畜牧兽医,2016,43(1):191-196. ZHANG X,SU Y,YANG S B,et al.Cloning of inhibin-α gene and its mRNA expression pattern in the ovary of Congjiang Xiang pig[J].China Animal Husbandry & Veterinary Medicine,2016,43(1):191-196.(in Chinese)
[2] SHENG J,YANG Y,LIU W,et al.Location of oocyte-specific linker histone in pig ovaries at different developmental stages postpartum[J].Theriogenology,2015,83(7):1203-1212.
[3] LAVOIE H A.Transcriptional control of genes mediating ovarian follicular growth,differentiation and steroidogenesis in pigs[J].Molecular Reproduction & Development,2017,84(9):788-801.
[4] 荀文娟,曹婷,周汉林,等.能量来源对五指山猪生长性能、初情期日龄及子宫和卵巢组织发育的影响[J].家畜生态学报,2017,38(6):41-45. XUN W J,CAO T,ZHOU H L,et al.Effects of different energy sources on growth performance,age of puberty,and uterus and ovarian development for Wuzhishan pigs[J].Journal of Domestic Animal Ecology,2017,38(6):41-45.(in Chinese)
[5] STOCCO D M.StAR protein and the regulation of steroid hormone biosynthesis[J].Annual Review of Physiology,2001,63(1):193-213.
[6] RAO R M,JO Y,BABBTARBOX M,et al.Regulation of steroid hormone biosynthesis in R2C and MA-10 Leydig tumor cells:Role of the cholesterol transfer proteins StAR and PBR[J].Endocrine Research,2002,28(4):387-394.
[7] HAAS B J,PAPANICOLAOU A,YASSOUR M,et al.De novo transcript sequence reconstruction from RNA-Seq using the Trinity platform for reference generation and analysis[J].Nature Protocols,2013,8(8):1494-1512.
[8] KIM D,PERTEA G,TRAPNELL C,et al.TopHat2:Accurate alignment of transcriptomes in the presence of insertions,deletions and gene fusions[J].Genome Biology,2013,14(4):R36.
[9] TRAPNELL C,WILLIAMS B A,PERTEA G,et al.Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation[J].Nature Biotechnology.2010,28(5):511-515.
[10] TRAPNELL C,WILLIAMS B A,PERTEA G,et al.Transcript assembly and abundance estimation from RNA-Seq reveals thousands of new transcripts and switching among isoforms[J].Nature Biotechnology,2010,28(5):511-515.
[11] 刘畅,王嘉福,黄世会,等.高低产群香猪X染色体的结构变异比较分析[J].农业生物技术学报,2017,25(7):1045-1058. LIU C,WANG J F,HUANG S H,et al.Comparative analysis of structural variations on chromosome X in two Xiang pig (Sus scrofa) groups with high and low litter size[J].Journal of Agricultural Biotechnology,2017,25(7):1045-1058.(in Chinese)
[12] 李丽莎,彭永东,郑晓宁,等.山羊PMEL基因启动子活性及转录调控元件分析[J].畜牧兽医学报,2017,48(5):826-835. LI L S,PENG Y D,ZHENG X N,et al.Analysis of the promoter activity and transcriptional regulatory elements of goat PMEL gene[J].Acta Veterinaria et Zootechnica Sinica,2017,48(5):826-835.(in Chinese)
[13] STIGTER J,SCHNEIDER M,BRUIJN F J.Azorhizobium caulinodans nitrogen fixation (nif/fix) gene regulation:Mutagenesis of the nifA -24/-12 promoter element,characterization of a ntrA(rpoN) gene,and derivation of a model[J].Molecular Plant-Microbe Interactions,1993,6(2):238-252.
[14] 张雄,张勇,陈祥,等.猪肌分化因子1基因启动子区多态性及生物信息学研究[J].中国畜牧兽医,2016,43(5):1308-1315. ZHANG X,ZHANG Y,CHEN X,et al.Study on polymorphisms and bioinformatics of MyoD1 gene promoter region in pig[J].China Animal Husbandry & Veterinary Medicine,2016,43(5):1308-1315.(in Chinese)
[15] 王彩云,潘登,张召议,等.类固醇激素合成调节因子在小鼠下丘脑和卵巢上的分布[J].中国兽医杂志,2017,53(2):29-32. WANG C Y,PAN D,ZHANG Z Y,et al.Expression of steroidogenic factor in mice ovary and brain[J].Chinese Journal of Veterinary Medicine,2017,53(2):29-32.(in Chinese)
[16] MAKOTO K,TAKASHI T,HJAMES M Q,et al.Characterization and expression of steroidogenic acute regulatory protein and MLN64 cDNAs in trout[J].Endocrinology,2002,143(6):2062-2070.
[17] TREMBLAY J J,VIGER R S.GATA factors differentially activate multiple gonadal promoters through conserved GATA regulatory elements[J].Endocrinology,2001,142(3):977-986.
[18] SHEA-EATON W K,TRINIDAD M J,LOPEZ D,et al.Sterol regulatory element binding protein-1a regulation of the steroidogenic acute regulatory protein gene[J].Endocrinology,2001,142(4):1525-1533.
[19] MANNA P R,WANG X J,STOCCO D M.Involvement of multiple transcription factors in the regulation of steroidogenic acute regulatory protein gene expression[J].Steroids,2003,68(14):1125-1134.
[20] MOE M,MEUWISSEN T,LIEN S,et al.Gene expression profiles in testis of pigs with extreme high and low levels of androstenone[J].BMC Genomics,2008,4(1):1-16.
[21] SHEN W,LI T,HU Y,et al.Common polymorphisms in the CYP1A1 and CYP11A1 genes and polycystic ovary syndrome risk:A meta-analysis and meta-regression[J].Archives of Gynecology & Obstetrics,2014,289(1):107-118.
[22] NEWELL-FUGATE A E,TAIBL J N,ALLOOSH M,et al.Effects of obesity and metabolic syndrome on steroidogenesis and folliculogenesis in the female ossabaw mini-pig[J].PLoS One,2015,10(6):e0128749.
[23] SCHANTON M,MAYMÓ J,PÉREZ-PÉREZA,et al.Sp1 transcription factor is a modulator of estradiol leptin induction in placental cells[J].Placenta,2017,57:152-162.

香猪卵巢StAR和CYP11A1基因的差异表达研究

Study on Differential Expression of StAR and CYP11A1 Genes in Xiang Pig Ovary

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