牦牛死亡结构域蛋白(FADD)基因克隆与生物信息学分析

死亡结构域蛋白(FADD)是在生物细胞里起到信号转导作用的一种蛋白,在胚胎发育、细胞凋亡过程中发挥重要的生物学活性。实验以成年母牦牛的卵巢RNA为模板,利用RT-PCR技术克隆牦牛FADD基因,并进行了生物信息学分析。结果表明:牦牛FADD基因CDS区长度630 bp,编码209个氨基酸,蛋白分子式C996H1632N300O307S7,整个蛋白质带负电荷,总共原子数量3 242;分子质量23.0 ku,半衰期30 h;理论等电点6.11;消光系数18 240;不稳定指数49.08,属于不稳定蛋白;脂肪系数104.64,平均亲水系数-0.168,预测FADD蛋白为亲水蛋白。系统进化树表明,牦牛与哺乳动物亲缘关系近,与鱼亲缘关系最远,符合物种进化规律。FADD蛋白质的二级结构α螺旋、自由卷曲、β-转角和延伸链,占靶蛋白的比例分别为71.29%、22.97%、3.83%和1.91%,与三级结构相符。牦牛FADD蛋白有13.0%位于细胞核、52.2%位于细胞质、8.7%位于细胞外、13.0%位于线粒体、4.3%位于高尔基体和8.7%位于细胞骨架。该研究成功克隆出牦牛FADD基因CDS区,为进一步研究其功能提供了一定理论依据。

Cloning and Bioinformatics Analysis of Yak FADD Gene

Fas-associated protein with death domain(FADD)is a protein that transduces signals in biological cell.It plays a crucial role in biological activity in embryonic development and apoptosis.Total RNA of yak ovary was used as the template to clone FADD gene using RT-PCR and analyze its bioinformatics characteristics.The results showed that the length of the FADD gene CDS region is 630 bp,encoding a 209 amino acid protein.The protein molecular formula is C996 H1632 N300 O307 S7.It is negative charge with 3 242 of the total number of atoms,23.0 ku of the molecular weight,30 h of the half-life,6.11 of theoretical isoelectric point,and 18 240 of extinction coefficient.It is an unstable protein with 49.08 of instability index.Its fat coefficient is 104.64.FADD protein is predicted as a hydrophilic protein with-0.168 of average hydrophilic coefficient.The phylogenetic tree showed that yak is closes to mammal and far to fish,which is consistent to species evolutionary law.The secondary structure is consists of alpha helix(71.29%),random coil(22.97%),beta turn(3.83%)and extended strand(1.91%),which is consistent with the tertiary structure analysis.Cell localization analysis indicated that yak FADD was in the nucleus(13.0%),cytoplasm(52.2%),extracellular(8.7%),mitochondrion(13%),Golgi(4.3%) and cytoskeleton(8.7%).This experiment cloned successfully CDS of yak FADD gene,which provided a theoretical basis for further study its function.