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

我们鉴定了马红细胞氨基酸转运蛋白和KCL联合转运蛋白的cDNA，以及马的几个氨基酸转运蛋白基因的EST。氨基酸转运蛋白和KCL共转运蛋白基因的聚合酶链式反应分析我们在马基因组DNA2中发现了12个氨基酸转运蛋白基因的EST序列：溶质载体家族1A4(SLC1A4)、SLC1A5、SLC3A1、SLC3A2、SLC7A5、SLC7A6、SLC7A7、SLC7A8、SLC7A9、SLC7A10、SLC7A11和SLC43A1，以及KCL共转运蛋白基因：SLC12A4。氨基酸转运蛋白和KCL共转运蛋白基因的定位利用荧光原位杂交技术，获得了SLC7A10：INRA638E6和SLC12A4：INRA125B7的马BAC克隆，分别定位于马染色体ECA3p13和ECA10p15。包括KLC共转运蛋白基因阳性的BAC克隆我们在马基因组DNA中发现了9个与SLC12A4基因座相近的EST。BAC克隆INRA125B7含有LCAT、PSMB1O和PSKH1，但不包括NFATc3、RCD-8、NUTF2、C16orf56、TSNAXIP1和RANBP10.4。氨基酸转运蛋白和KCL共转运蛋白基因的鉴定我们发现了7个转运蛋白基因的RT-PCR产物：SLC1A5、SLC3A1、SLC3A2、SLC7A5、SLC7A7、SLC7A8、SLC7A11、SLC43A1和SLC12A4。我们还检测了马SLC3A2、SLC7A7、SLC7A11、SLC43A1和SLC12A4的开放阅读框核苷酸序列，这些序列具有较高的转录水平。结论如下：未成熟马红细胞转录了6个氨基酸转运蛋白基因：SLC1A5、SLC3A2、SLC7A7、SLC7A8、SLC7A11和SLC43A1，以及一个KCL共转运蛋白基因：SL-12A4。马基因组包括12个氨基酸转运蛋白基因：SLC1A4、SLC1A5、SLC3A1、SLC3A2、SLC7A5、SLC7A6、SLC7A7、SLC7A8、SLC7A9、SLC7A10、SLC7A11和SLC43A1，以及一个KCL联合转运蛋白基因：SLC12A4。马基因组SLC7A10和SLC12A4基因座的邻近区域分别与人类基因组的同源区域具有同源性。

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.