Table 1.
Epithelial tissue expression and canine genome status of genes studied and their RT-PCR primers used for the semi-quantitative gene expression analysis in MDCK, IEC-6 and C2C12 cells.
| Genes | Intestine and kidneya | Canine genomeb | Primers for MDCK (forward and reverse, 5′–3′)b | Product (bp) | Primers for IEC-6 and C2C12 (forward & reverse, 5′–3′)c | Product (bp) |
|---|---|---|---|---|---|---|
| Primer pair for RT-PCR normalization gene | ||||||
| GAPDH | yes | yes | AGTCCATCTCCATCTTCCAG CGTCACGCCACATCTTCC |
381 | ATCCCATCACCATCTTCCAG GCCATCACGCCACAGTTTCC |
393 |
| Primer pairs for differentiation-related genes (identified through human microarray) | ||||||
| C2CD2L | yes | predicted | CACCTGGGAAGTGAGTTGG CGTTGGAAAGGTGGCTCTCC |
622-5 | ||
| EXOC2 | yes | predicted | TCCGCTTGGTATTGAGATTG TTTCCGTTCCATCCGCTTC |
296-9 | GCGAATCCTCTTGGCATTG CTTTTTCAGTTCCATCTGCTTC |
307 |
| MYLK | yes | predicted | CTCCAGTGTCAGGTGTCGTC GCTGCCATTCTCACTGGTC |
531 | ||
| MYO7B | yes | predicted | CGACATCCACTTCAACCCC CCTGGTGACAAACTCCCCTC |
502 | ACCATCCGCAATGACAACTC GGACAGGGATTGTGTGTTTG |
523 |
| SDCBP2 | yes | predicted | TCTGGAGGACCTGAAGATGG CAGCCCCACAAGAGACG |
403 | TCTGGAGGACCTGAAGATGG GCAAACCCACAAGGGATGC |
419-31 |
| Primer pairs for chosen KIF genes (identified through mouse microarray) | ||||||
| KIF1A | yes | yes | CCCCTTGATGTCCGAGTGTC GAGTAGAGCGTGTCGGTGAG |
780 | ATCAAAGATGGCGTCACCAG TCGTGTCTCTCGGTCTTTGG |
804 |
| KIF5A | yes | predicted | TGCGGGGAGACAAGTTCATC GCACCCGATTCTTGTCCTCA |
417 | TGCGGGGAGACAAGTTCATC CTTGACAAACGGCACCCG |
428 |
| KIF5B | yes | predicted | GCAAGCAGAAAACGATGCC GTTGCTCTCAGTTTGTGTGC |
427 | ||
| KIF12 | yes | predicted | GGAAGACCTACACCCTGACC GACTCCCAAACTCCACCACC |
276 | TACACTCTGACTGGACCTCC CTGTTCCACATAGAAGCCTC |
423 |
| KIF13A | yes | predicted | AAGTGATGCGAGTGACAGGG CCAGCACAGCGTTTCTTTCC |
969 | GGCGGAGGATGACTCTTC CACTGCTCCACCAGCCTG |
858 |
| KIF21A | yes | predicted | CCGAAGGAGAACCACCACTC CAGGAAATGGGTTGATTATGCCC |
524 | ||
| KIF27 | yes | predicted | CACTTTGTGGACTTGGCTG GTTAGTTCTTCGCTCAGTTG |
923-38 | CACTTTGTGGACTTGGCTG TGAAATCCTGTCTTGATGGG |
1084 |
| KIFAP3 | yes | predicted | TTTCCACCAAGCCACAAGAG GGAAATGAATGCTGCCCAAC |
411 | GCCACAAGAGATGTCATAATC GGAAATGAGTGCTGTCCAACIEC−6 AAATGCGTGTTGCCCCACC2C12 |
399-401 |
| Primer pairs for chosen RAB genes (identified through mouse microarray) | ||||||
| RAB1A | yes | yes | GCGACTCTGGGGTTGGAAAG TTGCTCCAGGACCCATTCGC |
489 | ||
| RAB2A | yes | yes | GGTTAGAAGATGCCCGCCAG GTGGCATTAGTAGCAGCGTG |
292 | ||
| RAB5A | yes | yes | AGCAACAAGACCCAACGGG AACACTGACTCCTGGTTGGC |
623 | AGCAACAAGACCCAACGGG AACACTGACTCCTGGTTGGC |
623 |
| RAB8A | yes | yes | TGGTCAAGAACGGTTTCGG TGGCGAGAGTGAAAAATGCG | 305 | ||
| RAB10 | yes | yes | CCAGGAGCGATTTCACACC TTGCTCTTCCAGCCTGTCAC |
393 | ||
| RAB15 | yes | predicted | GAAGACCATAGAGGTAGATG TCAGGTTTGCCCTCATCC |
462 | ||
| RAB19 | yes | predicted | GAACACGATTGGGGTGGAC CATCAGCGTGAAGACCTCG |
385 | ||
| RAB25 | yes | predicted | ATGGGGAACAGAACCGAGG TGGAGGTCACTCTTGTTGCC |
389 | ||
| RAB27A | yes | yes | GGCAAGAGAGGTTTCGTAGC TCCCCTTCTCCTTTGCTTC |
423 | GGCAAGAGAGGTTTCGTAGC ACCCCTTCTCCTTCTCCTC |
423 |
| RAB27B | yes | predicted | ACCTCACCAGTCAACAGAGC TCTCTGCTGGTTTTTCCCC |
342 | ACCTCACCAGTCAACAGAGC TCTCTGCTGGTTTTTCCCC |
342 |
a
The expression in the intestine and kidney is confirmed by experimental evidences from a variety of mammalian species listed in the UniGene database.
b
Genes belong to the “yes” category if the presence in canine genome has been experimentally validated. Otherwise, primer design was based on the predicted transcripts in the canine genome.
c
IEC6 and C2C12 primers were designed based on validated Rattus norvegicus or Mus musculus genes.