摘要
Wnt10b是Wnt家族中重要成员之一,绵羊Wnt10b基因尚属未知。本研究利用RT-PCR和RACE技术克隆了绵羊Wnt10b基因的全序列,并对该基因进行了比较全面的生物信息学分析。结果表明:绵羊Wnt10b基因序列全长2321 bp,包括1116 bp的编码区序列,编码391个氨基酸;其可变剪接体Wnt10b-AS由于CDS序列部分缺失编码166个氨基酸。同源性分析发现,绵羊Wnt10b基因核苷酸和氨基酸序列与其它哺乳动物同源性较高。氨基酸序列分析发现该蛋白为亲水性不稳定蛋白,相对分子量为43.0254 k Da。预测含有6个磷酸化位点,5个豆蔻酰化位点,二级结构以无规则卷曲和α螺旋为主。生物信息学分析表明Wnt10b及Wnt10-AS均为分泌型跨膜蛋白,主要存在细胞外基质中,通过与细胞膜上受体结合,调控下游靶基因的表达。Wnt10b-AS由于缺失多个翻译后修饰位点,可能通过与主转录本不同的作用机制发挥生物学功能。本研究为进一步探讨Wnt10b基因在绵羊毛囊生长发育中的功能和作用机制奠定了信息学基础。
Wnt10b is a member of Wnt family, has remained unknown among sheep. To predict the mechanisms of Wnt10 b in the regulation of hair follicle, we obtained the complete c DNA of Wnt10 b gene by using RT-PCR and RACE, and an alternative splicing（Wnt10b-AS） was identified accidentally. Protein was analyzed using bioinformatics tools. The results showed that the Wnt10 b was composed of 2321 nucleotides, including a single open reading frame（1 116 bp）, encoding 391 amino acids. The Wnt10b-AS in which 229 bp in exon 4 was spliced out, encoding 166 amino acids. Bioinformatics analysis revealed that that the Wnt10 b in sheep shared high identity with multiple mammalian species. Wnt10 b contained one N-glycosylation site, six phosphorylation sites and five myristoylation sites, while only4 of them remained in Wnt10b-AS. The secondary structure of Wnt10 b included Helix（36.32%）, Random coil（53.45%） and Extended strand（10.23%）. The matured proteins encoded by Wnt10 b and Wnt10b-AS were probably hydrophilic and extracellular matrix. However, loss of posttranslational modification sites indicated that the Wnt10b-AS gene were probably involved in different physiological functions and forms of cellular regulation.
出处
《中国畜牧杂志》
CAS
北大核心
2016年第7期11-15,共5页
Chinese Journal of Animal Science
基金
国家转基因重大专项"高产超细毛转基因羊新品种培育"(2014ZX08008-001
2013ZX08008-001)
