仙居鸡抑制素α亚基成熟区的cDNA克隆及其在卵泡中表达的研究  

cDNA Cloning and mRNA Expression Pattern in Follicles of the Mature Inhibin αSubunit from Xianju Chicken

作　　者：傅衍[1] 牛冬[1] 阮晖[2] 余旭平[1] 陈功[1] 何国庆[2] 杨培新[1] 

机构地区：[1]浙江大学动物科学学院,杭州310029 [2]浙江大学农业工程与食品科学学院,杭州310029

出　　处：《Acta Genetica Sinica》2001年第8期707-715,共9页

基　　金：国家自然科学基金(批准号39900106)&&

摘　　要：根据发表的鸡抑制素α亚基序列设计引物，用ＲＴ－ＰＣＲ技术从仙居鸡卵泡的颗粒细胞总ＲＮＡ中扩增出了抑制素α亚基成熟区序列，并进行了克隆和测序。结果显示，仙居鸡成熟α亚基是由１１３个氨基酸（ａａ）残基组成的蛋白质，具有１个糖基化位点和７个半胱氨酸残基，与发表的鸡和哺乳类相应序列对比，其核苷酸序列的同源性分别为９８％和６１．４％～６８．７％，其预测氨基酸序列的同源性分别为９７．３％和６４．６％～６９％，且所测鸡α亚基成熟区的糖基化位点及半胱氨酸残基的数目和位置与发表的鸡和哺乳类相似，说明该亚基的序列及结构在不同物种间具高度保守性，提示其可能具重要的生理功能。鸡各级卵泡中α亚基ｍＲＮＡ表达丰度的定量分析显示，从ＳＹＦ到Ｆ１中，随着卵泡的成熟α亚基的表达量降低，其在ＳＹＦ和Ｆ６～８中表达量最高，在ＬＷＦ中表达量很低，说明α亚基在卵泡的吸收、选择及优势化过程中起重要调节作用。The mature region of Xianju chicken inhibin α- subunit was amplified from the total RNA of follicle granulosa cells by RTWR using the primer pair designed according to the reported cDNA sequence of chicken inhibin α-subunit,and this fragment of α-subunit was cloned and sequenced subsequently. The results revealed that the mature α-subunit of Xianju chicken was a fragment of 113 amino acids containing one glycosylation site and seven cysteine residues. It was approximately 98% and 61.4%- 68.7% identical in nucleotide sequence, 97.3% and 64.6%- 69% similar in deduced amino acid sequence, respectively, in the mature region to the chicken and mammalian inhibin α-subunit cDNA clone. As for the mature chicken inhibin α-subunit,the number of potential glycosylation site and cysteine residues was the same, and their corresponding positions in the amino acid sequences were almost identical as compared to chicken and mammalian inhibin α-subunit,which indicated that the inhibin α-subunit was highly conserved among different species, implying an important role of inhibin α-subunit in various animals. The quantitative analysis of competitive RT-PCR for inhibin α-subunit revealed that the expression of α-subunit decreased with further follicle maturity from SYF to F1 follicle. The highest level of inhibin α-subunit mRNA was found in the SYF and F6-8 follicles, which indicated that inhibin α-subunit played an important role during the course of follicular recruitment,selection and dominance.

关 键 词：仙居鸡 抑制素α亚基 CDNA克隆 卵泡 定量竞争RT-PCR 

分 类 号：S831[农业科学—畜牧学]