CDNA Cloning And Spacr-timr Expression Analysis Of Cationic Amino Acid Transporters Gene In Shanbei White Cashmere Goat

This study was to explore the main expression location and developmental pattern of cationic amino acid transporter gene 4F2hc(SLC3A2), y+LAT1(SLC7A7), y+LAT2(SLC7A6), r BAT(SLC3A1) and CAT-1(SLC7A1)in Shanbei White Cashmere goat and to provide certain theoretical support for further research on mechanism of amino acid absorption and transport in ruminant intestinal and feed manufacturing according to the physiological characteristics of goat.Primers were designed based on the sequences of 4F2 hc, y+LAT1, y+LAT2, r BAT and CAT-1 c DNA in Ovis aries which were deposited in the Gen Bank Data Library. Reverse Transcription PCR(RT-PCR) was used to ampilfy the total or partial c DNA Coding sequences(CDS), then the sequence information was analyzed.The tissue distribution patterns of 4F2 hc, y+LAT1, y+LAT2, r BAT and CAT-1 m RNA were determined by Real-time Quantitative PCR(RT-QPCR) using the heart, liver, kidney, brain, muscle, skin, rumen, duodenum, jejunum, ileum and colon tissues in three 1-day-old Shanbei White Cashmere goat. The m RNA expression law of 4F2 hc, y+LAT1, y+LAT2, r BAT were investigated using duodenum, jejunum and ileum tissues collected from total 15 Shanbei White Cashmere goats at the age of day 1, month 6, 8, 10 and 12(3 in each age). The main results showed that as follows:1. The full-length CDS sequence of Shanbei White Cashmere goat 4F2 hc, y+LAT1, y+LAT2 and r BAT c DNA and partial-length c DNA sequence of CAT-1 were successfully cloned.The similiarities of goat 4F2 hc, y+LAT1, y+LAT2, r BAT and CAT-1 gene with that in Ovis aries and Bos taurus were all very high.2. Expression of 4F2 hc, y+LAT1, y+LAT2, r BAT and CAT-1 varied from tissues in 1-day-old Shanbei White Cashmere goat. Expression of 4F2 hc m RNA in skin was significant higher than that in other tissues of 1-day-old goats(P<0.05). It was highly expressed in jejunum besidies in skin. Expression of y+LAT1 m RNA in small intestine was significant higher than that in other tissues of 1-day-old goats(P<0.05). Expression of y+LAT2 m RNA in brain and ileum was significant higher than that in other tissues of 1-day-old goats(P<0.05). Expression of r BAT m RNA was very high in kidney and small intestine of 1-day-old goats. Expression of CAT-1 m RNA was very high in brain, then in skin, ileum and kidney.3. 4F2 hc m RNA abundance of small intestine in 1-day-old-goat was significant higher than that in the other ages(P<0.05). However, there was no significant difference in goats of the other ages(P>0.05). y+LAT1 m RNA abundance of small intestine decreased with age. y+LAT2 m RNA abundance of duodenum and jejunum peaked in 10-month-old goats and then dropped, while in ileum, it was significantly lower in 1-day-old goats than that in the other ages(P<0.05). r BAT m RNA abundance of duodenum in 1-day-old goats was significant higher than that in the other ages(P<0.05) and no significant differences were found in goats of the other ages(P>0.05). It was decreased significantly with age in jejunum(P<0.05), but there was no significant difference in goats of all ages(P>0.05). CAT-1 m RNA abundance of duodenum and jejunum in 1-day-old goats was significant lower than that in the other ages(P<0.05), but it was significant higher in 1-day-old goats than that in the other ages(P<0.05).Results indicate: Cationic amino acid transporters gene were highly conserved in the evolutionary process of ruminants. The m RNA expression of cationic amino acid transporters gene exists some differences in defferent tissues of goats and it was differentially regulated and distributed with the differences of developmental stages and segments of small intestine in goats.