DMA FRAGMENT ENCODING MIH/VIH RELATED PEPTIDE IN GIANT FRESHWATER PRAWN MACROBRACHIUM ROSENBERGII (DE MAN)

摘要

Three repeatable bands of 1200 bp, 1000 bp and 650 bp were obtained by PCR amplification of genomic DMA of Macrobrachium rosenbergii with homologous primers designed from M. rosenbergiicDNA sequence. These three fragments after elution were used as templates in separate polymerase chain reactions with nested primers. In nested PCR, only 1200 bp fragment yielded a single band of 490 bp indicating that the 1200 bp fragment includes the open reading frame as well as 5' UTR sequence of M. rosenbergiiMIH/VIH gene.The size of 1200 bp and 490 bp nested PCR fragment indicates presence of intron(s). Overall the PCR based strategy for MIH/VIH amplification and cloning seems to be difficult due to high degree of sequence homology between various classes of neuropeptides and clusteral arrangement of neuropeptide encoding genes. The 1200 bp and the 490 bp MIH/VIH encoding fragments have been cloned and could be used as probes in genomic library screening and RFLP to tap MIH/VIH gene variation in the population.