Clay Latonya A, Wang Shiao Y, Wolters William R, Peterson Brian C, Waldbieser Geoffrey C
USDA, ARS, Catfish Genetics Research Unit, Thad Cochran National Warmwater Aquaculture Center, 141 Experiment Station Road, Stoneville, MS 38776, USA.
Biochim Biophys Acta. 2005 Dec 20;1731(3):139-48. doi: 10.1016/j.bbaexp.2005.10.001. Epub 2005 Oct 27.
The insulin-like growth factor I (IGF-I) gene was characterized in channel catfish. Partial cDNA sequence, missing exon 1 and part of exon 2, was obtained in 5'- and 3'-RACE experiments. Direct sequencing of two bacterial artificial chromosome clones revealed gene structure and provided sequence from 640 bp upstream of the initiator methionine to 136 bp beyond the polyadenylation site. Genomic sequence contained a putative TATA box 506 bp upstream of the initiator methionine. The 477-bp reading frame within five exons encoded a 159-amino acid (aa) pre-propeptide highly similar to IGF-I in higher vertebrates. The sequence encoding the signal peptide was unique in catfish and contained 70% G+C content with the potential for a stable stem-loop structure. Full-length cDNA was only maintained in recombination-deficient (DH10B) strain E. coli. Levels of IGF-I mRNA were highest in liver, followed by brain and muscle, then heart and kidney (P<0.05). A CT/GA dinucleotide microsatellite in intron 1 was highly polymorphic in commercial channel catfish, and permitted placement of the IGF-I gene on the catfish genetic map. However, specific IGF-I alleles were not correlated with differences in growth rate from 100 to 130 days post-hatch in USDA103 line catfish.
对斑点叉尾鮰的胰岛素样生长因子I(IGF-I)基因进行了表征。在5'-和3'-RACE实验中获得了缺失外显子1和部分外显子2的部分cDNA序列。对两个细菌人工染色体克隆进行直接测序,揭示了基因结构,并提供了从起始甲硫氨酸上游640 bp到多聚腺苷酸化位点下游136 bp的序列。基因组序列在起始甲硫氨酸上游506 bp处包含一个推定的TATA框。五个外显子内477 bp的阅读框编码了一个159个氨基酸(aa)的前原肽,与高等脊椎动物的IGF-I高度相似。编码信号肽的序列在斑点叉尾鮰中是独特的,G+C含量为70%,具有形成稳定茎环结构的潜力。全长cDNA仅在重组缺陷型(DH10B)大肠杆菌菌株中得以保留。IGF-I mRNA水平在肝脏中最高,其次是脑和肌肉,然后是心脏和肾脏(P<0.05)。内含子1中的CT/GA二核苷酸微卫星在商业斑点叉尾鮰中具有高度多态性,并允许将IGF-I基因定位在斑点叉尾鮰遗传图谱上。然而,特定的IGF-I等位基因与美国农业部103系斑点叉尾鮰孵化后100至130天的生长速率差异无关。