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

Yi Chuan. 2008 Jun;30(6):747-54. doi: 10.3724/sp.j.1005.2008.00747.
[Article in Chinese]

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.

MeSH terms

  • Adaptor Proteins, Signal Transducing / chemistry*
  • Adaptor Proteins, Signal Transducing / genetics
  • Adaptor Proteins, Signal Transducing / metabolism*
  • Amino Acid Sequence
  • Animals
  • B-Cell CLL-Lymphoma 10 Protein
  • Base Sequence
  • Cell Line
  • Cloning, Molecular / methods*
  • Gene Expression Profiling / methods*
  • Humans
  • Immunity, Innate / genetics*
  • Kidney / metabolism
  • Liver / metabolism
  • Molecular Sequence Data
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment
  • Spleen / metabolism
  • Swine

Substances

  • Adaptor Proteins, Signal Transducing
  • B-Cell CLL-Lymphoma 10 Protein
  • BCL10 protein, human

Associated data

  • GENBANK/EU088132