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

本研究鉴定了马红细胞氨基酸转运蛋白和KCl共转运蛋白的cDNA，以及马中几个氨基酸转运蛋白基因的EST.氨基酸转运蛋白基因和氯化钾协同转运蛋白基因的PCR分析在马基因组DNA中，我们发现了12个氨基酸转运蛋白基因的EST：溶质转运蛋白家族1A 4（SLC 1A 4）、SLC 1A 5、SLC 3A 1、SLC 3A 2、SLC 7A 5、SLC 7A 6、SLC 7A 7、SLC 7A 8、SLC 7A 9、SLC 7A 10、SLC 7A 11和SLC 43 A1，以及一个氯化钾协同转运蛋白基因：SLC 12 A4.氨基酸转运蛋白和KCL协同转运蛋白基因的定位获得了SLC 7A 10：INRA 638 E6和SLC 12 A4：INRA 125 B7的马BAC克隆，并通过荧光原位杂交（FISH）将其分别定位于马染色体ECA 3 p13和ECA 10 p15上. KLC共转运蛋白基因阳性BAC克隆的包含基因我们在马基因组DNA中发现了9个与SLC 12 A4位点邻近的EST。BAC克隆INRA 125 B7含有LCAT、PSMB 10和PSKH 1，但不含NFATC 3、RCD-8、NUTF 2、C16 orf 56、TSNAXIP 1和RANBP 10.4。氨基酸转运蛋白和KCL协同转运蛋白基因cDNA的鉴定我们发现了7个转运蛋白基因的RT-PCR产物：SLC 1A 5、SLC 3A 1、SLC 3A 2、SLC 7A 5、SLC 7A 7、SLC 7A 8、SLC 7A 11、SLC 43 A1和SLC 12 A4。我们还检测到了马的SLC 3A 2、SLC 7A 7、SLC 7A 11、SLC 43 A1和SLC 12 A4的开放阅读框的核苷酸序列，这些基因具有较高的转录水平。结论如下：未成熟马红系细胞转录6个氨基酸转运蛋白基因：SLC 1A 5、SLC 3A 2、SLC 7A 7、SLC 7A 8、SLC 7A 11和SLC 43 A1，以及1个KCL协同转运蛋白基因：SL-12 A4。马基因组包括12个氨基酸转运蛋白基因：SLC 1A 4、SLC 1A 5、SLC 3A 1、SLC 3A 2、SLC 7A 5、SLC 7A 6、SLC 7A 7、SLC 7A 8、SLC 7A 9、SLC 7A 10、SLC 7A 11和SLC 43 A1，以及KCL共转运蛋白基因：SLC 12 A4。马基因组的SLC 7A 10和SLC 12 A4位点的邻近区域分别与人类基因组的同源区域同线性。

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.