肉鸡MTHFR基因的克隆和表达水平研究

发布时间：2018-05-26 23:29

  本文选题：肉鸡 + MTHFR基因　； 参考：《浙江农业学报》2016年12期

【摘要】：采用RT-PCR方法克隆了鸡MTHFR(methylenetetrahydrofolate reductase,MTHFR)基因,利用生物信息学方法预测蛋白结构,应用定量PCR(q PCR)技术检测组织表达特性。结果表明,鸡MTHFR基因的c DNA序列长度为2 342 bp(Gen Bank登录号:KU351685),开放阅读框长1 956 bp,编码651个氨基酸;进化树分析表明,鸡MTHFR氨基酸序列与绿头鸭和鸿雁的关系较近。MTHFR氨基酸序列包含保守的MTHFR superfamily结构域,为亲水蛋白;二级结构主要是α-螺旋(38.71%)和无规则卷曲(36.41%);亚细胞定位显示,该蛋白存在于细胞质和细胞核的概率分别为60.9%和30.4%。q PCR分析表明,MTHFR基因在所检测鸡的8种组织中均有表达,其中在心脏中表达水平最高,在腿肌中的表达水平最低;叶酸缺乏试验表明,母鸡叶酸缺乏能极显著增加子代肉鸡MTHFR基因在肝脏中的表达水平(P0.01)。研究结果可为进一步研究MTHFR基因的结构和功能以及叶酸在家禽生产中的应用提供资料。
[Abstract]:The chicken MTHFR (methylenetetrahydrofolate reductase, MTHFR) gene was cloned by RT-PCR method. The protein structure was predicted by bioinformatics and quantitative PCR (Q PCR) was used to detect the tissue expression characteristics. The results showed that the C DNA sequence of the chicken MTHFR gene was 2342 BP, and the length of the open reading frame was 1956. The evolution tree analysis showed that the relationship between the MTHFR amino acid sequence of the chicken MTHFR and the green goose duck and the wild goose was closer to the.MTHFR amino acid sequence including the conservative MTHFR superfamily domain and the hydrophilic protein; the two grade structure was mainly alpha helix (38.71%) and the irregular curl (36.41%), and the subcellular location showed that the protein was found in the cytoplasm and in the cytoplasm. The probability of 60.9% and 30.4%.q PCR analysis showed that the MTHFR gene was expressed in the 8 tissues of the chickens detected, among which the expression level in the heart was the highest and the expression level in the leg muscles was the lowest. The folate deficiency test showed that the maternal folate deficiency could significantly increase the expression level of the MTHFR gene in the liver of the broilers (P0 .01). The results can provide information for further research on the structure and function of MTHFR gene and the application of folic acid in poultry production.
【作者单位】： 临沂大学生命科学学院;
【基金】：国家自然科学基金项目(31372333) 山东省自然科学基金项目(ZR2013CL012) 临沂大学国家级大学生创新创业训练计划项目(201510452021)
【分类号】：S831
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本文编号：1939409