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CRABP-ICellular Retinoic Acid-Binding Protein Type I
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Cloning of bovine CRABP-I and -II genes by reverse transcriptase-PCR (RT-PCR)
Moreover, bovine CRABP-I cDNA was amplified from adipose tissue using RT-PCR with a specific primer for bovine CRABP-I (forward, 5'-TGCCACCATGCCCAACTT-3; and reverse, 5'-CGCCTTCATTCCCGAACA-3).
Comparative real-time PCR was performed using a Roter Gene 2000 system (Cobett Research, Australia) and a QuantiTect SYBR Green RT-PCR kit (QIAGEN, Germany) according to the manufacturers' instructions, with the following primer pairs: CRABP-I sense, 5'-AAGGCTTTGAGGAGGAGACC-3'; CRABP-I antisense, 5'-TCAGGATGAGTTCGTCGTTG-3'; CRABP-II sense, 5'-AAACAGGAGGGGGACACTTT-3'; CRABP-II antisense, 5'-CCCATTTCACCAGGCTCTTA-3'; 18S rRNA sense, 5'-CCCGAAGCGTTTACTTTGAA-3'; and 18S rRNA antisense, 5'-CCCTCTTAATCATGGCCTCA-3'.
The amino-acid sequence comparison presented in Figure 2A reveals an overall degree of identity between bovine CRABP-I and -II of approximately 75%.