Molecular cloning, recombinant expression, and growth-promoting effect of mud carp (Cirrhinus molitorella) insulin-like growth factor-I

Gen Comp Endocrinol. 2006 Sep 1;148(2):203-12. doi: 10.1016/j.ygcen.2006.03.014. Epub 2006 May 16.

Abstract

A full-length cDNA encoding insulin-like growth factor-I (IGF-I) was cloned from mud carp (Cirrhinus molitorella) liver tissue using reverse transcription polymerase-chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. The IGF-I precursor cDNA consists of 822 bp in size with a 218 bp 5'-untranslated region and 118 bp 3'-untranslated region. The 486 bp open reading frame encodes a 161 amino acid peptide with a molecular weight of 17.9 kDa. The deduced IGF-I amino acid sequence shared 82.5-97% and 82.5-84% sequence identity with fish and mammalian counterparts, respectively. The mature IGF-I was overexpressed in Escherichia coli, and the expression level of recombinant mcIGF-I reached to 34.1% of the cell total protein. After purification and refolding of recombinant mcIGF-I, growth-promoting effect of recombinant mcIGF-I was investigated, the results showed that the recombinant mcIGF-I significantly enhanced the growth rate of juvenile tilapia. After 6-week treatment, the growth rates of group 1 and 2 were 53 and 67.3% higher than the saline-treated control group. The recombinant mcIGF-I was more effective than recombinant mcGH to enhance the growth rate of juvenile tilapia. The recombinant mcIGF-I-treated fish revealed no significant changes of content of protein, lipid, ash and moisture in muscle.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Body Weight / drug effects
  • Carps / genetics*
  • Carps / metabolism*
  • Cloning, Molecular
  • DNA, Complementary / isolation & purification
  • Escherichia coli / metabolism
  • Fish Products / analysis
  • Growth and Development / drug effects*
  • Insulin-Like Growth Factor I / genetics*
  • Insulin-Like Growth Factor I / isolation & purification
  • Insulin-Like Growth Factor I / metabolism*
  • Insulin-Like Growth Factor I / pharmacology*
  • Molecular Sequence Data
  • Muscles / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism*
  • Recombinant Proteins / pharmacology*
  • Sequence Homology, Amino Acid
  • Tilapia / growth & development

Substances

  • DNA, Complementary
  • Recombinant Proteins
  • Insulin-Like Growth Factor I