Difference between revisions of "Kathryn"
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'''Notes 1/12/16''' | '''Notes 1/12/16''' | ||
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Only .1 g of organ taken: possibility that connective tissue was taken, not representative of entire organ possibly | Only .1 g of organ taken: possibility that connective tissue was taken, not representative of entire organ possibly | ||
total RNA > mRNA using beads that attach to polyA tails of mRNA | total RNA > mRNA using beads that attach to polyA tails of mRNA | ||
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mRNA > CDNA: using reverse transcriptase, dNTP, use primers that has every possibly combination of 6 nucleotides so all mRNA is transcribed | mRNA > CDNA: using reverse transcriptase, dNTP, use primers that has every possibly combination of 6 nucleotides so all mRNA is transcribed | ||
CDNA: has been transcribed as mRNA and then changed into DNA to make more stable form | CDNA: has been transcribed as mRNA and then changed into DNA to make more stable form | ||
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| + | ---- | ||
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| + | '''Research''' | ||
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| + | The molecular correlate of system B0, the major apical neutral amino acid transporter in kidney and intestine, is B0AT1 (SLC6A19) (46), a protein of 634 amino acids. Currently, no splice variants of the transporter have been reported. The human SLC6A19 shows very little activity in heterologous expression systems; hence, the mouse transporter has been characterized in more detail. In agreement with functional studies, B0AT1 transports all neutral amino acids, albeit to a varying extent. Vmax values appear to be fairly similar, but affinities are different for each amino acid. The order of preference is Met = Leu = Ile = Val > Gln = Asn = Phe = Cys = Ala > Ser = Gly = Tyr = Thr = His = Pro > Trp > Lys. This order is in partial agreement with studies in the intestine (280). The transporter shows some affinity for lysine; | ||
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| + | In situ hybridization and immunocytochemical analysis showed that the transporter is expressed in the kidney proximal convoluted tubule (46, 297) and in all parts of the small intestine but not in the colon (Fig. 1). Expression increases from the duodenum to the ileum (297, 373). The transporter is confined to the apical membrane. The signal was more intense towards the tip of the villi | ||
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| + | - http://physrev.physiology.org/content/88/1/249#sec-45 | ||
Revision as of 19:38, 14 January 2016
THIS IS KATHRYN'S PAGE
Notes 1/12/16
Only .1 g of organ taken: possibility that connective tissue was taken, not representative of entire organ possibly total RNA > mRNA using beads that attach to polyA tails of mRNA - randomly fragment mRNA (since you can only read from an end to 75 base pairs) now get a lot more accurate reads, now know more about entire sequence mRNA > CDNA: using reverse transcriptase, dNTP, use primers that has every possibly combination of 6 nucleotides so all mRNA is transcribed CDNA: has been transcribed as mRNA and then changed into DNA to make more stable form
Research
The molecular correlate of system B0, the major apical neutral amino acid transporter in kidney and intestine, is B0AT1 (SLC6A19) (46), a protein of 634 amino acids. Currently, no splice variants of the transporter have been reported. The human SLC6A19 shows very little activity in heterologous expression systems; hence, the mouse transporter has been characterized in more detail. In agreement with functional studies, B0AT1 transports all neutral amino acids, albeit to a varying extent. Vmax values appear to be fairly similar, but affinities are different for each amino acid. The order of preference is Met = Leu = Ile = Val > Gln = Asn = Phe = Cys = Ala > Ser = Gly = Tyr = Thr = His = Pro > Trp > Lys. This order is in partial agreement with studies in the intestine (280). The transporter shows some affinity for lysine;
In situ hybridization and immunocytochemical analysis showed that the transporter is expressed in the kidney proximal convoluted tubule (46, 297) and in all parts of the small intestine but not in the colon (Fig. 1). Expression increases from the duodenum to the ileum (297, 373). The transporter is confined to the apical membrane. The signal was more intense towards the tip of the villi
