农业现代化研究2011，Vol.32Issue(6)：756-760,5.
藏猪兴奋性氨基酸转运载体EAA C1基因的克隆与原核表达
Gene Cloning and Prokaryotic Expressing of Excitatory Amino Acid Carrier 1 in Tibetan Pig
摘要
Abstract
Excitatory amino acid carrier 1 (EAAC1) is the major transporter of glutamate in the intestine. A cDNA fragment about 1600 bp was amplified from the total RNA of jejunum in Tibetan piglets by RT-PCR with a pair of specific primers based on the sequences of human. The recombinant vector was constructed by inserting the cDNA fragment into the prokaryotic expression vector pET-32a (+) and then transformed into E. Coli BL21 (DE3). Sequence analysis showed that the ORF of EAAC1 cDNA in Tibetan piglets was 1575 bp in length encoding 524 amino acid residues with a molecular mass of 57 kD and isoionic point of 5.34. Its GenBank accession number is GQ375513. Three putative extracellular N-glycosylation sites, eight potential protein kinase C phosphorylation sites and one deduced cAMP/cGMP-dependent protein kinase phosphorylation site were identified. Seven proposed transmembrane domains and a N-terminus signal peptide were presented in EAAC1 sequence. The SDS-PAGE displayed that the expressed protein was consistent with the size of expected protein. These findings maybe provide some scientific basis for the study on function and regulation of EAA C1 in pigs.关键词
藏猪/兴奋性氨基酸转运载体/序列分析/原核表达Key words
Tibetan pig/excitatory amino acid carrier 1/sequence analysis/prokaryotic expression分类
生物科学引用本文复制引用
傅德智,孔祥峰,杨焕胜,褚武英,李铁军,印遇龙..藏猪兴奋性氨基酸转运载体EAA C1基因的克隆与原核表达[J].农业现代化研究,2011,32(6):756-760,5.基金项目
国家自然科学基金项目(编号:31110103909、30928018、30901040、31001016) （编号:31110103909、30928018、30901040、31001016）
中国科学院知识创新工程重要方向项目(编号:KZCX2-EW-412). （编号:KZCX2-EW-412）
