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Analysis of equine amino acid permease and KCL co-transporter genes

马氨基酸通透酶和 KCL 协同转运蛋白基因分析

基本信息

批准号：
15580266
负责人：
HANZAWA Megumu
金额：
$ 2.05万
依托单位：
Tokyo University of Agriculture
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

We report the identification of cDNAs for amino acid transporters and KCL co-transporter of equine erythrocytes, and EST for several amino acid transporter genes in the horse.1. PCR analysis of amino acid transporter and KCL co-transporter genesWe identified twelve ESTs for amino acid transporter gene : Solute Carrier Family 1A4 (SLC1A4), SLC1A5, SLC3A1, SLC3A2, SLC7A5, SLC7A6, SLC7A7, SLC7A8, SLC7A9, SLC7A10, SLC7A11 and SLC43A1, and a KCL co-transporter gene : SLC12A4 in the horse genome DNA.2. Mapping of amino acid transporter and KCL co-transporter genesEquine BAC clones were obtained for SLC7A10 : INRA638E6 and SLC12A4 : INRA125B7, and mapped to equine chromosomes ECA3p13 and ECA10p15, respectively, by fluorescence in situ hybridization (FISH).3. Included genes of the KLC co-transporter gene positive BAC cloneWe identified nine EST close to the SLC12A4 locus in the horse genome DNA. The BAC clone INRA125B7 contained LCAT, PSMB1O and PSKH1, but not include NFATC3, RCD-8, NUTF2, C16orf56, TSNAXIP1 and RANBP10.4. Identification of cDNA for amino acid transporter and KCL co-transporter genesWe found RT-PCR products for seven transporter genes : SLC1A5, SLC3A1, SLC3A2, SLC7A5, SLC7A7, SLC7A8, SLC7A11, SLC43A1 and SLC12A4. We detected also nucleotide sequences for open reading frame of equine SLC3A2, SLC7A7, SLC7A11, SLC43A1 and SLC12A4, which had much transcription levels. These sequence data coded functional transporters.The conclusions are settled as follows.Immature equine erythroid cell transcribe six amino acid transporter genes : SLC1A5, SLC3A2, SLC7A7, SLC7A8, SLC7A11 and SLC43A1, and a KCL co-transporter gene : SL-12A4. The horse genome include twelve amino acid transporter genes : SLC1A4, SLC1A5, SLC3A1, SLC3A2, SLC7A5, SLC7A6, SLC7A7, SLC7A8, SLC7A9, SLC7A10, SLC7A11 and SLC43A1 and a KCL co-transporter gene : SLC12A4. The vicinity areas of SLC7A10 and SLC12A4 locus of horse genome have synteny to homologous area of human genome respectively.

我们报道了马红细胞中氨基酸转运蛋白和KCL共转运蛋白的cdna鉴定，以及马中几种氨基酸转运蛋白基因的EST鉴定。氨基酸转运基因和KCL共转运基因的PCR分析我们鉴定了12个氨基酸转运基因的ESTs：溶质载体家族1A4 （SLC1A4）、SLC1A5、SLC3A1、SLC3A2、SLC7A5、SLC7A6、SLC7A7、SLC7A8、SLC7A9、SLC7A10、SLC7A11和SLC43A1，以及马基因组dna中的KCL共转运基因SLC12A4。3.氨基酸转运体和KCL共转运体基因定位利用荧光原位杂交（FISH）技术，获得了SLC7A10: INRA638E6和SLC12A4: INRA125B7的马BAC克隆，并分别定位到马染色体ECA3p13和ECA10p15上。包括KLC共转运基因阳性BAC克隆的基因我们在马基因组DNA中鉴定出9个靠近SLC12A4位点的EST。BAC克隆INRA125B7含有LCAT、psmb10和PSKH1，但不含NFATC3、RCD-8、NUTF2、C16orf56、tsnaxp1和RANBP10.4。氨基酸转运基因和KCL共转运基因cDNA的鉴定我们发现了7个转运基因的RT-PCR产物：SLC1A5、SLC3A1、SLC3A2、SLC7A5、SLC7A7、SLC7A8、SLC7A11、SLC43A1和SLC12A4。我们还检测到马的SLC3A2、SLC7A7、SLC7A11、SLC43A1和SLC12A4的开放阅读框核苷酸序列，这些序列的转录水平较高。这些序列数据编码功能转运蛋白。结论如下：未成熟马红细胞转录6个氨基酸转运基因：SLC1A5、SLC3A2、SLC7A7、SLC7A8、SLC7A11和SLC43A1，以及一个KCL共转运基因：SL-12A4。马基因组包括12个氨基酸转运基因：SLC1A4、SLC1A5、SLC3A1、SLC3A2、SLC7A5、SLC7A6、SLC7A7、SLC7A8、SLC7A9、SLC7A10、SLC7A11和SLC43A1，以及一个KCL共转运基因SLC12A4。马基因组SLC7A10位点和SLC12A4位点邻近区域分别与人类基因组同源区域具有同源性。