Construction of full-length cDNA library with goose muscular tissue and sequence analysis of anUbI gene

The muscular tissue of breast was dissected from Jilin White goose at 8 weeks of age in the present study.The long-distance PCR method was used to amplify double-strand cDNA based on the SMART techniques for construction of a full-length cDNA library.After digestion with restriction endonuclease SfiⅠ,a modified vector of pBluescript II SK plasmid with the adaptors containing SfiⅠSfiⅠA and SfiⅠB sites was used to recombine with the cDNA products amplified.The recombinants were cloned by transformation into competent Escherichia coli DH2α.A plasmid cDNA library with goose muscle was constructed.The results showed that the titer of the cDNA library is 1.01×106 pfu /ml,in which the percentage of recombinant clones is 97%.The length of most cDNA inserted was between 0.25 kb and 1.1 kb identified by gel electrophoresis after cDNA PCR amplification.The unigene ratio was 66.7% and the percentage of complete cDNA sequences was 80% by estimating from the 24 clones sequenced randomly.A full-length anUbⅠcDNA was found by aligned with those of its homologues in the NCBI database using the BlastN program,and the accession number in GenBank is DQ420616.The sequence length of coding region of anUbⅠcDNA is 918 bp,it encodes 305 amino acids consisting of the polyubiquitin protein.By comparison,high similarities of 87.3-93.7% in the coding region sequences of polyubuquitin genes were found in 16 species including chicken.It is helpful to study muscle development of goose at molecular level in the future.