畜牧兽医学报 ›› 2007, Vol. 38 ›› Issue (2): 110-114.doi:

• 遗传繁育 • 上一篇    下一篇

西农萨能羊两个泌乳阶段差异表达基因SAA3的克隆和序列分析

武会娟,罗军,张丽娟,韩雪峰,杨宝进,王海滨,单翠燕,张宁,余刚   

  1. 西北农林科技大学动物科技学院,杨凌712100
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-02-25 发布日期:2007-02-25
  • 通讯作者: 武会娟

Cloning and Sequence Analysis of SAA3 Gene Differentially Expressed in Mammary Gland at Two Lactation Stages of Xinong Saanen Goat

WU Hui-juan, LUO Jun , ZHANG Li-juan, HAN Xue-feng, YANG Bao-jin, WANG Hai-bin, SHAN Cui-yan, ZHANG Ning, YU Gang   

  1. College of Animal Science and Technology , Northwest A&F University, Yangling 712100, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2007-02-25 Published:2007-02-25

摘要: 为了研究不同泌乳期乳腺组织中差异表达基因,利用抑制性削减杂交技术构建西农萨能羊泌乳60d乳腺组织及泌乳28d乳腺组织中差异表达基因文库,用Real-Time PCR技术验证阳性克隆血清淀粉样蛋白A3(SAA3)基因,通过RT-PCR方法克隆西农萨能羊乳腺组织SAA3,并进行序列比对和功能预测。结果成功构建了西农萨能羊不同泌乳期乳腺组织中差异表达基因文库,筛选克隆了乳腺组织SAA3基因,GenBank登录号为:DQ839400,编码区长度为396bp,含有131个氨基酸。西农萨能羊乳腺组织中SAA3与牛(GenBank:NM_181016)、兔(GenBank:M64696.1)、人(GenBank:BC020795)、鼠(GenBank:NM_011315)核苷酸同源性分别为95%、84.3%、81.3%和81.9%,氨基酸同源性为93%、76%、72%、72%;在编码区261-287位较人、兔、鼠SAA编码区多27个碱基,连续大于5个氨基酸的保守区域有6个,较人SAA多3个潜在功能基序。

Abstract: To discover genes differently expressed between lactation periods, suppression subtractive hybridization method was employed to construct a cDNA library expressed in mammary gland of 60^th but not or little in 28^th. Serum amyloid A protein was one of the positive clones which was verified by Real-time PCR. Moreover the gene was cloned by RT-PCR and the sequence was analyzed. Results showed that a differently expressed library was successfully created in mammary gland of 60^th lactation. The goat mammary gland SAA3 complementary DNA has an open reading frame of 396 nucleotides encoding a putative protein of 131 amino acids, the GenBank number was DQ839400. The homologies of nucleotide and peptide sequence of Xinong Saanen goat mammary gland SAA3 with bovine(NM_181016), rabbit (M64696. 1), human(BCA)20795) and mouse's(NM_011315) are 95%, 84. 3%, 81.3% and 81.9% and 93%, 76%, 72% and 72% respectively. The 27 nucleotides in goat SAA3 261-287 were deleted in that of human, rabbit, and mouse. There were six conserved domains longer than 5 amino acids in the amino acids sequences among them. Goat SAA3 had three additional potential binding motifs compared to human's.