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中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (08): 38-44    DOI:
    
Cloning,Sequencing Analysis and Prokaryotic Expression of the IGF-Ⅰ Gene from Mink
RONG Min1, YANG Fu-He1, HU Chang-Zhong2, GENG Xiu-Mei1, WANG Gui-Wu1, WU Qiong1
1. The Institute of Special Economic Animals and Plants, Chinese Academy of Agricultural Sciences,Jilin132109,China
2. Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing100094,China
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Abstract  

The sequence of mink IGF-Ⅰgene was obtained and analyzed,and the fusion protein of the IGF-Ⅰ was produced in E.coli. By RTPCR,liver IGF-Ⅰfrom mink were isolated. The IGF-ⅠcDNAs encoding 153 amino acids,and showed high degree of homology(>90%)with other reported sequences(gold monkey,lesser panda,giant panda,A.tiger,horse,et al.) at nucleotide and amino acids levels.Two differences in mink amino acids sequences were found from sequence alignment.Whether these differences affect the molecular conformation or function for the growth factor's processing or these changes in the molecular structure may be responsible for the abnormal ability of growth and reproductive by influencing the function of IGF-Ⅰ have not yet been determined.The DNA fragment encoding the mature peptide of mink IGF-Ⅰwere subcloned to the pGEX-6P-1 expression vector and highly expressed in E.coli.BL21 with IPTG induction. The expression fusion proteins(pGEX-6P-1 IGF-Ⅰ)were mostly existed in soluble form and were about 34kDa.The induced cells culture were proved to be active IGF-Ⅰantigens. The mink IGF-Ⅰ gene was successfully cloned and expressed.Furthermore,the structure and function of this gene were analyzed and predicted. These result provided a good basis for further study of its biological activity.



Key wordsMink       Insulin-like growth factor-Ⅰ Clone Prokaryotic expression     
Received: 09 March 2009      Published: 28 July 2009
Cite this article:

RONG Min, YANG Fu-He, HU Chang-Zhong, GENG Xiu-Mei, WANG Gui-Wu, TUN Qiong. Cloning,Sequencing Analysis and Prokaryotic Expression of the IGF-Ⅰ Gene from Mink. China Biotechnology, 2009, 29(08): 38-44.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I08/38

[1] 刘阳.蓝狐GDF8基因cDNA克隆及GDF8基因与IGFⅠ基因表达研究[学位论文].东北林业大学.2005
Liu Y.A study on cDNA clone of GDF8 gene and the mRNA expression of GDF8 gene and IGF1 gene in blue fox.Harbin:Northeast Forestry University,2005
[2]  Daughaday W H,Hall K,Salmon W D,et al.J Clin Endocrinol Metab,1987,65:1075~1076
[3]  Froesch E R,Schmid C,Schwander J,et al.Am Rev Physiol,1985,47:443~467
[4]  Cohik W S, Clemmons D R. The insulin2like growth factors Ⅰ.Annu  Rev Physiol, 1993, 55: 131
[5] Jones J I,Clemmons D R.Insulinlike growth factors and their binding proteins:biological actions.Endocr Rev,1995,16:3~34
[6] Upton Z,Yandell C A,Degger B G,et al.Evolution of insulinlike growth factorⅠ(IGFⅠ)action:in vitro characterization of vertebrate IGFⅠproteins.Comp Biochem Physiol B Biochem Mol Biol,1998,121:35~41
[7] Lund P K,Moatsstaasts B M,Hynes M A,et al.Somatomedin C/insulinlike growth factor Ⅰand insulinlike growth factorⅡ mRNAs in rat fetal and adult tissues.J Biol Chem,1986,261:14539~14544
[8] Von Heijne G. A new method for predicting signal sequence cleavage sites.Nucleic Acids Res,1986,14:4683~4690
[9] Hodgson D R,May F E,Westley B R,et al.Involvement of phenylalanine 23 in the binding of IGFⅠ to the insulin and typeⅠIGF receptor.Regul Pept,1996,66(3):191~196
[10] Cascieri M A,Chicchi G G,Applebaum J J,et al.Identification of the domain of IGFⅠresponsible for high affinity binding to the types 1 and 2 IGF receptors(IGFR1, IGFR2),insulin receptor(IR)and binding proteins(BP).FASEB J.1988,2:8577.
[11] Nagagawa S H,Tger H S.Role of the COOHterminal Bchain domain in insulinreceptor interactions;identification of perturbations involving the insulin main chain.J Biol Chem,262(25):12054~12058
[12] Bayne M L,Applebaum J,Chicchi G G,et al.The roles of tyrosines 24,31 and 60 in the high affinity binding of insulinlike growth factorⅠto the type 1 insulinlike growth factor receptor.J Biol Chem,1990,265:15648~15652
[13]  胡细连.大熊猫等濒危动物IGFⅠ基因的克隆、表达与组织分布研究[学位论文] .浙江大学,2005
Hu X L.Molecular Cloning,Expression and Tissue Distribution of Insulinlike Growth Factor Ⅰ in Endangered Animals.Hangzhou:Zhejiang University,2005
[14]  Stephen S,Ditchkoff Leon J Spicer,Ronald E,et al.Concentrations of insulinlike growth factorⅠin adult male whitetailed:associations with serum testosterone,morphometrics and age during and after the breeding season.Comparative Biothemistry and Physiology,2001, Part A 129:887~895
[15] Moriyama S,Duguay S J,Conlon J M,et al. Recombinant coho salm on insulinlike growth factor Ⅰexpression in Escherichia coli,purification and characterization.Eur J Biochem,1993,218:205~211
[16]  华益民,林浩然,等.草鱼IGFⅠcDNA的克隆和在原核生物中的表达.动物学报,2001,47(3):274~279
Hua Y M,Lin H R.Cloning and expression in prokaryote of grass carp IGF Cdna.Acta Zoologica Sinica,2001,47(3):274~279
[17]  张建峰.鸡IGFⅠ对成骨细胞功能的影响及其基因多态性与蛋鸡骨代谢相关性的研究[学位论文].南京农业大学.2005
Zhang J F.Studies on effects of chicken IGFI on osteoblastic functions and its correlation of gene polymorphism to bone metabolism in cage layers.Nanjing:Nanjing Agricultural University,2005

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