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| 版纳微型猪近交系雄激素受体AR基因的克隆和表达分析 |
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王配1,2, 李罗刚1,2, 张霞1,2, 王淑燕1,2, 霍海龙3, 曾日彬1,2, 刘民2, 王雪飞2, 霍金龙1,2
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| (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 教务处, 昆明 650031) |
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| 摘要: |
| 为探讨猪雄激素受体（Androgen receptor，AR）基因的序列、结构、表达和功能，以版纳微型猪近交系（Banna mini-pig inbred line，BMI）睾丸组织为材料，用RT-PCR技术获得猪AR基因cDNA序列，进行生物信息学分析，应用荧光定量技术明确其mRNA的多组织表达特征。结果表明：BMI AR基因cDNA长3 257 bp（GenBank登录号：KU705631），包含2 688 bp的全长编码区，位于猪X染色体，含有9个外显子和8个内含子；推导出的BMI AR蛋白含有1个核定位信号、1个DBD结构域和1个LBD结构域，无信号肽，无跨膜螺旋；与黄牛、山羊、人、猩猩、大鼠和小鼠的AR蛋白依次有94.2%、94.0%、90.2%、89.0%、88.1%和87.5%的同源性；其二级结构中以无规则卷曲和α螺旋为主，延伸链和β转角只在局部出现；在二级结构预测结果的基础上利用同源建模构建了其三级结构；AR基因mRNA在被检测的BMI 15个组织中呈现差异表达现象，表达量最高的组织为睾丸。该研究结果为进一步研究AR基因在BMI中的生理功能及调节机制提供了依据。 |
| 关键词: 雄激素受体 版纳微型猪近交系 组织表达谱 生物信息学 |
| DOI：10.11841/j.issn.1007-4333.2018.01.12 |
| 投稿时间：2017-01-20 |
| 基金项目:国家自然科学基金项目（31160439，31460580，31660650，31660637）；云南省应用基础研究计划项目（2016FB046，2016FD027） |
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| Cloning and expression analysis of androgen receptor (AR) gene in Banna mini-pig inbred line |
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WANG Pei1,2, LI Luogang1,2, ZHANG Xia1,2, WANG Shuyan1,2, HUO Hailong3, ZENG Ribin1,2, LIU Min2, WANG Xuefei2, HUO Jinlong1,2
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| (1.Key Laboratory of Banna Mini-pig Inbred Line of Yunnan Province, Yunnan Agricultural University, Kunming 650201, China;2.Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China;3.Teaching Affairs Department, Yunnan Vocational and Technical College of Agriculture, Kunming 650031, China) |
| Abstract: |
| In order to explore sequence and structure of pig androgen receptor (AR), the cDNA of AR gene was cloned from the testis of Banna mini-pig inbred line (BMI) by RT-PCR for sequencing and characterization of AR was analyzed by bioinformatics analysis. The expression pattern of AR in different tissues was detected by qPCR. A cDNA of 3 257 bp (GenBank accession number:KU705631) of BMI AR was obtained, which contained 2 688 bp opening reading frame. Genome structure analysis showed it localized to Sus scrofa chromosomes X and was consisted of nine exons and eight introns. Functional bioinformatics analysis indicated that BMI AR protein contained one nuclear location signal, one DBD domain, one LBD domain, no transmembrane region and no signal peptide. The predicted amino acid sequence of this gene had high homology with the cattle (94.2%), goat (94.0%), human (90.2%), chimpanzee (89.0%), rat (88.1%) and mouse (87.5%). The secondary structure of BMI AR protein was mainly consisted of random coils and α-helices, β turns and extended strands were locally present. Its tertiary structure was also predicted by homology modeling based on the secondary structure data. AR mRNA was differentially expressed in the 15 tissues of BMI with the highest expression level in testis. This research provided foundations for further study on the physiological function and regulatory mechanisms of AR gene in BMI. |
| Key words: androgen receptor Banna mini-pig inbred line tissue expression patterns bioinformatics |
