| 摘要: |
| 为分析鸡印度刺猬因子(Indian hedgehog，IHH)基因生物信息学及IHH基因在不同组织中特异性表达规律，选取体重相近、健康强健的6只鸡，借助生物信息学手段，对IHH基因结构、启动子和CpG岛进行分析，并通过实时荧光定量PCR检测IHH基因在鸡心、肝、脾、肺、肾、软骨、下丘脑、胸肌、十二指肠和胸腺组织中的特异性表达规律，同时对IHH蛋白进行相似性比较，并利用生物信息学分析IHH蛋白的理化和结构特性。结果表明，1)鸡IHH基因开放阅读框长度为1 227 bp，可编码408个氨基酸，具有3个外显子、4个核心启动子区和1个CpG岛。2)成功扩增IHH基因，IHH基因在鸡各组织中均有不同水平的mRNA表达，在软骨组织中相对表达量最高，在心、脾和胸腺组织中的表达量相对较低。3)鸡IHH蛋白在禽类中保守性较高，IHH蛋白大小为44.829 36 ku，属于偏碱性蛋白，具有1个跨膜端和1个信号肽；IHH蛋白二级结构中无规则卷曲的比例最高(44.12%)，只含有1个 HH-Signal功能域，内含大部分丝氨酸磷酸化位点，该蛋白还有14个苏氨酸磷酸化位点和5个酪氨酸磷酸化位点，鸡IHH蛋白存在1处N型糖基化位点和6处O型糖基化位点。综上，鸡IHH基因有3个外显子，有多个启动子区，在软骨组织中极显著高表达，鸡IHH蛋白是具有偏碱性、强热稳性和低亲水性的分泌型蛋白，IHH氨基酸序列在物种进化过程中比较保守，本研究为深入研究鸡IHH基因对软骨发育的调控作用提供依据。 |
| 关键词: 鸡 IHH 生物信息学 特异性表达 |
| DOI：10.11841/j.issn.1007-4333.2023.10.11 |
| 投稿时间：2022-10-10 |
| 基金项目:国家自然科学基金项目(31802028)；江苏省家禽遗传育种重点实验室开放课题(JQLAB-KF-202101)；高校优秀青年人才支持重点项目(gxyqZD2022017)；安徽省教育厅高校自然科学研究重点项目(2022AH050928)；教育部产学研合作协同育人项目(221000488095409)；2021年省级继续教育教学改革重点项目(2021jxjy026) |
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| Bioinformatics and expression pattern analysis of chicken IHH gene |
|
JIN Sihua,SHUI Fei,XU Yuan,JIA Yuqing,XIA Jingjing,WANG Xin,JIA Fumin,GENG Zhaoyu*
|
| (College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China) |
| Abstract: |
| In order to analyze the bioinformatics of chicken Indian hedgehog(IHH)gene and the specific expression of IHH gene in different tissues, a total of six healthy and strong chickens with similar weight were selected. The structure, promoter and CpG island of IHH gene were analyzed by bioinformatics, and the specific expression of IHH gene in chicken heart, liver, spleen, lung, kidney, cartilage, hypothalamus, pectoral muscle, duodenum and thymus was detected by quantitative real-time PCR(qRT-PCR). At the same time, the similarity of IHH protein was compared. The physicochemical and structural characteristics of IHH protein were analyzed by bioinformatics. The results showed as follows: 1)The open reading frame of chicken IHH gene was 1 227 bp in length, which can encode 408 amino acids, and had 3 exons, 4 core promoter regions, and 1 CpG island. 2)IHH gene was were successfully amplified. IHH gene had different levels of expression in chicken tissues, with the highest relative expression in cartilage tissue and relatively low expression in heart, spleen and thymus tissue. 3)Chicken IHH protein was highly conserved in poultry. The size of IHH protein was 44. 829 36 ku, which belonged to alkaline protein with one transmembrane terminal and one signal peptide. The proportion of random coils in the secondary structure of IHH protein was the highest(44. 12%), containing only one HH-Signal domain and most of the serine phosphorylation sites. In addition, the protein also had 14 threonine phosphorylation sites and 5 tyrosine phosphorylation sites. There were one N-type glycosylation and six O-type glycosylation sites in IHH protein. In summary, chicken IHH gene has three exons and multiple promoter regions, which are highly expressed in cartilage tissue. Chicken IHH protein is a secreted protein with alkaline, strong thermal stability and low hydrophilicity. The amino acid sequence of IHH is relatively conserved during species evolution. These results provide a basis for further study of chicken IHH gene regulation of cartilage development. |
| Key words: Chicken IHH bioinformatics analysis specific expression |
