Isolation of DNA Sequence Flanking T-DNA by Thermal Asymmetric Interlaced PCR from Monascus Pigment-producing Mutants

Abstract:

In this paper，thermal asymmetric interlaced PCR(TAIL-PCR) was adopted to amplify DNA sequences flanking T-DNA of pigment-producing mutants of Monascus spp. Seven DNA fragment which renged from 500bp to 1300bp were isolated. Then the fragment were sequenced and their functions were analyzed with Blast tool on line supplied by NCBI. The results showed that one of the DNA sequences was of similarity to the nucleotide sequence was of similarity to the necleotide sequence coding for the developmental regulator of flbA from Aspergillus fumigatus Af293 and the similarity of them amounted to 82%. The successful amplification to Monascus spp.by TAIL-PCR provides an efficient method to isolate DNA sequences flanking T-DNA from the monascus insertional mutants on a large scale and will be beneficial to investigate the functional gene of Monascus spp.

Key words:Monascus spp.，T-DNA insertion，Mutants，TAIL-PCR