Abstract:
In order to study the effect of
IGF-Ⅰ gene on growth traits of topmouth culter (
Culter alburnus Basilewsky),
IGF-Ⅰ gene was cloned from the genomic DNA of topmouth culter in this study. The gene spanned 14567 bp and comprised five exons and five introns. The five exons of
IGF-Ⅰ were 298, 160, 182, 36, and 1360 bp long, respectively. The hypothetical open reading frame of the
IGF-Ⅰ cDNA precursor was 486 bp, encoding a putative protein of 161 amino acids. The precursor peptide included a signal peptide (44 amino acids), a mature peptide (70 amino acids), and an E peptide (47 amino acids). The mature peptide comprised four regions: B, C, A, and D. The A and B domains were conserved with six cysteine residues in these two regions to form three disulfide bonds. The B region contained a conserved
IGF-Ⅰ receptor recognition sequence (Phe
B23-Tyr
B24-Phe
B25). Analysis of the E peptide showed that
IGF-Ⅰ of the topmouth culter was an Ea-2 type. The
IGF-Ⅰ amino acid sequence of the topmouth culter had high sequence identity with
IGF-Ⅰ proteins from Cyprinidae fish, ranging from 94 to 100%. Phylogenetic analysis of
IGF-Ⅰ amino acid sequences showed that
IGF-Ⅰ of the topmouth culter does not cluster with the fish from subfamily Cultrinae.
IGF-Ⅰ is expressed in all 10 tested tissues with the highest mRNA level in liver, modest level in spleen, heart, testis, and brain, and low level in kidney, gill, stomach, and ovary. The lengths of the four introns of topmouth culter
IGF-Ⅰ were 1170, 9364, 251, and 1746 bp, respectively. The variation in the sequence of the introns among species was greater than that of the exons with the highest variation in the third intron. Six microsatellite loci were found in the introns of
IGF-Ⅰ with (GATG)
5AATAT (ATAG)
11 in intron 1, and (CT)
8, (TTA)
5, (AC)
13, (TG)
12, and (ATT)
5 in intron 2. Four of 6 microsatellite loci were polymorphic, and the genotypes of the four microsatellite loci had no significant correlation with the growth traits of 120 topmouth culter cultured in the same pond (
P>0.05). These results provide molecular basis for studying the expression pattern, function and transcriptional regulation of the
IGF-Ⅰ gene.