Molecular cloning of ACRP30 (adipocyte complement-related protein of 30 kDa) gene and its impacts on glucose and lipid metabolism in orange-spotted grouper (epinephelus coioides)

• Main Authors: Chia-Fang Lee, 李佳芳
• Other Authors: Jenn-Kan Lu
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/11202566782324360906

碩士 === 國立臺灣海洋大學 === 水產養殖學系 === 93 === Adipose tissue plays an active role in monitoring and controlling whole-body metabolism by secreting a variety of bioactive molecules collectively termed adipokines. ACRP30 (Adipocyte Complement-Related Protein of 30 kDa) belongs to adipokines, which can regulates glucose and lipid homeostasis. In the present study, we have isolated the coding region of rabbit (Oryctolagus cuniculus) ACRP30 cDNA which fragment containing 735 bp encoded for 245 amino acids and a partial ACRP30 cDNA isolated from adipose tissue of orange-spotted grouper (Epinephelus coioides) containing 339bp encoded for 113 amino acids. Comparison of the identity of rabbit cDNA with various mammal ACRP30 sequence is ranging from 73％ to 84％, and the similarity of rabbit ACRP30 amino acid is ranging from 81％ to 84％. The grouper ACRP30 partial sequence is very similar to other vertebrates ACRP30, the identity of nucleotide sequence is ranging from 83％ to 99％, and the similarity of grouper amino acid is ranging from 90％ to 99％. RT-PCR analysis demonstrated that grouper ACRP30 mRNA is mainly expressed in adipose tissue. Expression of ACRP30 mRNA was first detected in yolk sac stage of grouper embryo, suggests that ACRP30 may play an important role in the metabolism of nutrients during yolk sac absorption stage. Expression of ACRP30 gene increased in 0~2h after scheduled feeding time. Feeding with high lipid diet could decrease ACRP30 mRNA expression. The ACRP30 mRNA decreased after starvation, and increased in restriction feeding of high lipid diet. An ACRP30 inducer, thiazolidinedione (TZD), can induce ACRP30 mRNA expression in adipose tissue of grouper and resulted in lipid level in liver and plasma glucose and triglycerides level decreased. Employment of E. coli expression system to produce recombinant globular domain ACRP30 protein of grouper, the molecular weight of rACRP30 is about 13 kDa, and it will be used for improvement glucose and lipid metabolism in finfish.