Abstract:

Total RNA was isolated from Nile tilapia pituitary gland. Then we cloned the cDAN encoding growth hormone from the the Nile tilapia pituitary by Reverse Transcription-PolymeraseChain Reaction(RT-PCR), sequence of 785bp.The result of sequence analysis and restriction map shows that an open reading frame of tilapia growth hormone gene contains 615 base pairs which code for a polypeptide of 204 amino acids including 17 amino acids of the signal peptlde and 187 amino acids of the mature growth hormone. Nucleotide sequence and amino acid sequence of tilapia growth hormone gene are as same as Rentier-Delrue’s tilapia GH cDNA in code region.The objective fragment was inserted into the downstream of the carrier promoter and combined with the report gene EGFP.The combinant plasmid was verified by PCR of the bacterium liquid，restriction enzyme digestion and gene sequencing.The pEGFP-GH recombinant is transfected into 293T cells by lipofection respectively. The results showed that the expressing fluorescent cells are more than 90% by pEGFP-N1; but the pEGFP-GH is less weaker intensity. The phenomena show that the Nile tilapia GH gene has good activity in eukaryotic cells.