cDNA Library Construction from Antler Cartilage of the Red Deer (Cervus elaphus) and Cloning Insulin-like Growth Factor II Gene
LIU Xue-dong, ZHANG Zi-dong, ZHENG Dong, MA Jian-zhang (Animal Genetics and Molecular Biology Laboratory, College of Wildlife Resources, Northeast Forestry University, Harbin 150040)
【Objective】 To investigate molecular mechanisms of antler cartilage growth and development at the functional genomic level and further clarify function of the Insulin-like growth factorⅡ (IGF2) gene. 【Method】 The SMART technique was used to construct a cDNA library from antler cartilage of the red deer (Cervus elaphus) and conserved primers were further designed to screen cDNA library and isolate IGF2 gene. 【Result】 The primary cDNA library contained 1.8×106 independent clones, 89.8% of which were recombinant; titer of the amplified library was 1.4×1010 pfu·ml-1 and average length of exogenous inserts was 1.2 kb. Bioinformatic analysis based on IGF2 genes of pig, horse, cattle, sheep, human, mouse and rat showed CDS region of the red deer IGF2 gene (GenBank accession No: EF177491) was 540 bp, which encoded a 179-amino-acids-long peptide. Compared with other seven species, IGF2 of the red deer had no tryptophan, and contained the most histidine and threonine. Moreover, phylogenetic tree reconstructed disclosed the highest similarities among IGF2 of deer, cattle and sheep. 【Conclusion】 The results demonstrated that the reliability and representativeness of the cDNA library were constructed, which would provide a further platform for studying regeneration mechanisms of antler cartilage of the red deer.