Construction of a BAC library and identification of Dmrt1 gene of the rice field eel, Monopterus albus

Journal Article · Fri Sep 22 04:00:00 EDT 2006 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/j.bbrc.2006.07.145· OSTI ID:20854479

Songhun, Jang ^[1]; Fang, Zhou ^[1]; Laixin, Xia ^[1]; Wei, Zhao ^[1]; Hanhua, Cheng ^[1]; Rongjia, Zhou ^[1]

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1. Department of Genetics and Center for Developmental Biology, College of Life Sciences, Wuhan University, Wuhan 430072 (China)

A bacterial artificial chromosome (BAC) library was constructed using nuclear DNA from the rice field eel (Monopterus albus). The BAC library consists of a total of 33,000 clones with an average insert size of 115 kb. Based on the rice field eel haploid genome size of 600 Mb, the BAC library is estimated to contain approximately 6.3 genome equivalents and represents 99.8% of the genome of the rice field eel. This is first BAC library constructed from this species. To estimate the possibility of isolating a specific clone, high-density colony hybridization-based library screening was performed using Dmrt1 cDNA of the rice field eel as a probe. Both library screening and PCR identification results revealed three positive BAC clones which were overlapped, and formed a contig covering the Dmrt1 gene of 195 kb. By sequence comparisons with the Dmrt1 cDNA and sequencing of first four intron-exon junctions, Dmrt1 gene of the rice field eel was predicted to contain four introns and five exons. The sizes of first and second intron are 1.5 and 2.6 kb, respectively, and the sizes of last two introns were predicted to be about 20 kb. The Dmrt1 gene structure was conserved in evolution. These results also indicate that the BAC library is a useful resource for BAC contig construction and molecular isolation of functional genes.

• OSTI ID: 20854479
• Journal Information: Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 2 Vol. 348; ISSN BBRCA9; ISSN 0006-291X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
CHROMOSOMES
DNA
EEL
EXONS
GENES
HYBRIDIZATION
POLYMERASE CHAIN REACTION
RICE
SCREENING