摘要
为了解苏淮猪BMP4基因的序列特征和表达特征,本研究采用克隆测序技术获得苏淮猪BMP4基因编码区全序列,利用生物信息学方法分析BMP4基因的序列特征,采用RT-PCR技术检测BMP4基因在苏淮猪不同组织中的表达情况,构建苏淮猪BMP4基因真核表达载体并利用Western blot技术检测BMP4真核表达载体的表达情况。结果显示苏淮猪BMP4基因编码区序列长度为1 230 bp,编码409个氨基酸,与其他哺乳动物氨基酸序列的一致性均在97%以上;苏淮猪BMP4编码蛋白含有典型的TGF-β前肽和TGF-β样结构域;BMP4基因在苏淮猪卵巢组织中高表达;成功构建苏淮猪BMP4基因真核表达质粒pc DNA3.1-BMP4,转染后可显著上调猪卵巢颗粒细胞BMP4蛋白表达水平。
The coding sequence of Suhuai pig BMP4 gene obtained by cloning and sequencing was analyzed by bioinformatics and the expression patterns were identified by RT-PCR. The full length coding region of BMP4 was 1 230 bp in length,encoding a protein with 409 amino acids which shared more than 97% similarities with other mammals. BMP4 protein of Suhuai pig contained TGF-β propeptide and TGF-β domain. RT-PCR assays showed that BMP4 is widely expressed in tissues and organs. BMP4 protein level was upregulated in porcine ovarian granulosa cells transfected with recombinant plasmid pc DNA3. 1-BMP4. The results laid a foundation for investigation on the role of BMP4 in porcine follicular development.
出处
《江苏农业学报》
CSCD
北大核心
2016年第3期581-587,共7页
Jiangsu Journal of Agricultural Sciences
基金
江苏省农业科技自主创新基金项目[SCX(12)2097]
关键词
苏淮猪
BMP4
表达模式
真核表达
Suhuai swine
BMP4
expression pattern
eukaryotic expression
