不同肌肉特异性启动子IGF2表达载体构建及对牛骨骼肌卫星细胞增殖的影响

doi:10.11843/j.issn.0366-6964.2015.04.007

Abstract

Abstract:

The aim of this study was to construct IGF2 expression vector with efficient and specific promoter and study its effects on bovine skeletal muscle satellite cell proliferation.The 3 eukaryotic expression vectors containing IGF2 and muscle-specific promoter were transfected into bovine skeletal muscle satellite cells and fetal fibroblasts，the relative expression of IGF2 was detected by RT-PCR to screen efficient and specific vector.EdU experiments were carried out and RT-PCR was used to detect the cell cycle-related genes.The different promoter （desminpro，CMV-MyoGpro and MyoGpro-double） IGF2 expression vectors were constructed successfully.The test results of RT-PCR showed that pGL3-desminpro-IGF2 was an efficient and specific vector.The vector transfected into cultured cell in vitro increased the speed of cell proliferation and up-regulated the expression of cell cycle-related genes，CDK6 and IGF2.The result has laid an important foundation for the production of transgenic bovine.

Key words: IGF2, muscle-specific promoter, skeletal muscle satellite cells, cell proliferation

CLC Number:

S823
S813.3

SUN Xiao-li，LI Shu-feng，TONG Hui-li，ZHANG Wei-wei，YIN Hong-yan，YAN Yun-qin. The Construction of Different Muscle-specific Promoter IGF2 Expression Vector and Its Impact on Bovine Skeletal Muscle Satellite Cell Proliferation[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(4): 555-560.

References

［1］MICKE G C，SULLIVAN T M，MCMILLEN I C，et al.Protein intake during gestation affects postnatal bovine skeletal muscle growth and relative expression of IGF1，IGF1R，IGF2 and IGF2R［J］.Mol Cell Endocrinol，2011，332(1)：234-241.
［2］朱婵，徐勇飞.IGF2 的生物学特性及其与疾病关系的研究进展［J］.国际检验医学杂志，2010，(9)：966-969.
ZHU C，XU Y F.Biological characteristics of IGF2 and Research progress of its relationship with disease ［J］.International Journal of Laboratory Medicine，2010，(9)：966-969.(in Chinese)
［3］GERRARD D E，OKAMURA C S，RANALLETTA M A，et al.Developmental expression and location of IGF-I and IGF-II mRNA and protein in skeletal muscle［J］.J Anim Sci，1998，76(4)：1004-1011.
［4］彭凤兰.胰岛素样生长因子 2 (Igf2) 研究进展［J］.实用预防医学，2008，14(6)：1963-1966.
PENG F L.The research progress of insulin-like growth factor2 (Igf2) ［J］.Practical Preventive Medicine，2008，14(6)：1963-1966.(in Chinese)
［5］CONSTÂNCIA M，HEMBERGER M，HUGHES J，et al.Placental-specific IGF-II is a major modulator of placental and fetal growth［J］.Nature，2002，417(6892)：945-948.
［6］DECHIARA T M，ROBERTSON E J，EFSTRATIADIS A.Parental imprinting of the mouse insulin-like growth factor II gene［J］.Cell，1991，64(4)：849-859.
［7］HAYASHI S，ASO H，WATANABE K，et al.Sequence of IGF-I，IGFⅡ，and HGF expression in regenerating skeletal muscle ［J］.Histochem Cell Biol，2004，122(5):427-434.
［8］袁建龙，王玮，金永，等.人工合成启动子特异启动 IGF-1真核载体的构建与绵羊成纤维细胞转染［J］.内蒙古大学学报：自然科学版，2010，41(6)：678-682.
YUAN J L，WANG W，JIN Y，et al.The construction of synthetic promoter promote IGF-1 eukaryotic vector and transfected into sheep fibroblasts ［J］.Inner Mongolia University：Natural Science，2010，41(6)：678-682.(in Chinese)
［9］LEVINOVITZ A，JENNISCHE E，OLDFORS A，et al.Activation of insulin-like growth factor II expression during skeletal muscle regeneration in the rat：correlation with myotube formation［J］.Mol Endocrinol (Baltimore，Md.)，1992，6(8)：1227-1234.
［10］李聪，马凯丽，刘增力，等.IGFs 及其信号通路研究进展［J］.生物技术通报，2013，(9)：43-46.
LI C，MA K L，LIU Z L，et al.The research progress of IGFs and its signaling pathways ［J］.Biotechnology Bulletin，2013，(9)：43-46.(in Chinese)
［11］DUPONT J，KARAS M，LEROITH D.The potentiation of estrogen on insulin-like growth factor I action in MCF-7 human breast cancer cells includes cell cycle components［J］. J Biol Chem，2000，275(46)：35893-35901.
［12］杜巍，李树峰，佟慧丽，等.牛结蛋白基因启动子的克隆及功能的初步分析［J］.畜牧兽医学报，2014，45(1)：56-61.
DU W，LI S F，THONG H L，et al.Cloning and functional analysis of bovine desmin gene promoter［J］. Acta Veterinaria et Zootechnica Sinica，2014，45(1)：56-61.(in Chinese)
［13］WANG X，LU M，FENG L H，et al.Effects of CMV enhancer on activity and specificity of bovine MyoG gene promoter［J］.J Northeast Agric University，2013，20(4)：34-38.
［14］王秋华，曹允考，李树峰，等.牛MyoG基因启动子的克隆及功能的初步分析［J］.畜牧兽医学报，2012，43(1)：37-43.
WANG Q H，CAO Y K，LI S F，et al.Cloning and functional analysis of bovine MyoG gene promoter［J］.Acta Veterinaria et Zootechnica Sinica，2012，43(1)：37-43.(in Chinese)
［15］DUPONT J，PIERRE A，FROMENT P，et al.The insulin-like growth factor axis in cell cycle progression［J］.Horm Metab Res，2003，35(11/12)：740-750.
［16］董雅静.IGF2 真核表达载体的构建及绵羊骨骼肌细胞转基因研究［D］.杨凌：西北农林科技大学，2012.
DONG Y J.The construction of IGF2 eukaryotic vector and the gene transfer in sheep skeletal muscle cells［D］.Yangling：Northwest A&F University，2012.(in Chinese)

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The Construction of Different Muscle-specific Promoter IGF2 Expression Vector and Its Impact on Bovine Skeletal Muscle Satellite Cell Proliferation

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