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HEREDITAS ›› 2008, Vol. 30 ›› Issue (6): 747-754.doi: 10.3724/SP.J.1005.2008.00747

• 研究报告 • Previous Articles     Next Articles

In silico identification, molecular cloning and verification of a novel pig gene homologous to human BCL10 of in-nate immunity and its preliminary expression profiles in pigs

HE Jian-Feng; ZHANG Yan-Ming; DUAN Hui-Juan; CAO Jin-Suo; ZHANG De-Li   

  1. Division of Virology, Immunology & Bioinformatics, Department of Preventive Veterinary Medicine, College of Veterinary Medicine , Northwest A & F University, Yangling 712100, China
  • Received:2007-12-25 Revised:2008-02-25 Online:2008-06-10 Published:2008-06-10
  • Contact: ZHANG De-Li

Abstract:

We identified and characterized a novel swine gene Bcl10 with GenBank (Accession No. EU088132) which was homologous to human BCL10 (B-cell CLL/lymphoma 10) gene. The full-length cDNA of 925 bp for swine BCL10 was in silico cloned, and then its ORF of 702 bp coding 233 amino acid residues was verified by RT-PCR and DNA sequencing. NCBI BLAST assay indicated that this cDNA contained three extrons with a length of 57, 289 and 356 bp respectively, and it was located on the chromosome 4 of pig genome. Using semi-quantitative PCR, preliminary expression profiles of swine BCL10 were verified in different swine tissues. The expression of swine BCL10 was verified by GFP marker and RT-PCR assay. We found that, BCL10 expressed in high level in swine spleen, and with a modest level in thymus, brain and lymph node; low level mRNA was expressed in liver and not detectable level in kidney. The swine BCL10 gene was cloned to the GFP-containing eukaryotic expression vector pEGFP-C1 and transfected to PK-15 cell line by lipofectin. BCL10 was ex-pressed effectively in PK-15 cells. In summary, we cloned a novel swine BCL10 gene, and investigated its expression char-acters. This will be the fundament of the future research on its function.