牡蛎胰岛素相关多肽3克隆表达及纯化

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Abstract Objective To clone and construct a BL21 expressing strain containing insulin related polypeptide 3 (IRP3) gene, and isolate, purifiy, identifiy and renaturat the expressed protein. Methods IRP3-pET30a positive strain BL21 was cloned by DNA recombination technique, the plasmid IRP3-PET30A was successfully constructed by agarose gel electrophoresis after double enzyme digestion, and IRP3/HIS purified protein was obtained by nickel affinity chromatography, SDS-PAGE and Western blot were further used to confirm the purification effect of IRP3/HIS fusion protein. Results Plasmid IRP3-PET30A was successfully cloned and BL21 expressing strain containing plasmid IRP3-pET30a was screened, purified IRP3/HIS fusion protein was obtained from the supernatant of inclusion body lysis and renaturated by dialysis. Conclusion The successful cloning of IRP3 gene and the expression and purification of IRP3/HIS fusion protein lay a foundation for the preparation of IRP3 protein antibody and the application of IRP3 protein as in vitro growth factor additive. What we have done will surely be useful to the antibody preparation for the IRP3 protein and the application of growth factor additive in vitro.

Key words： Insulin-related polypeptides 3 Genetic cloning Expression Purification

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	YANG Bingye LI Ying CHEN Zhongwei▲

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YANG Bingye LI Ying CHEN Zhongwei▲. Cloning expression and purification of insulin-related polypeptides 3 from Oyster#br#[J]. 中国医药导报, 2021, 18(30): 19-22.

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https://www.yiyaodaobao.com.cn/EN/ OR https://www.yiyaodaobao.com.cn/EN/Y2021/V18/I30/19

[1] 张惠婷.海水养殖贝类加工利用研究进展及展望[J].福建轻纺，2021（1）：43-45.
[2] 蔡水淋，陈晓婷，刘淑集，等.牡蛎肉的综合开发研究进展[J].渔业研究，2020，42（5）：527-532.
[3] 孙为刚.海蛎子[J].走向世界，2020（52）：80-81.
[4] 蒋如浩，慕永通.中国牡蛎养殖业集聚特征分析[J].中国渔业经济，2021，39（1）：55-61.
[5] 邹琰，张天文，刘广斌，等.太平洋牡蛎人工育苗技术[J].科学养鱼，2020（10）：62.
[6] 曾志南，宁岳.福建牡蛎养殖业的现状、问题与对策[J].海洋科学，2011，35（9）：112-118.
[7] 林丹，孙敏秋，张克烽，等.福建牡蛎产业发展形势分析[J].中国水产，2019（3）：53-57.
[8] 张立娟，何毛贤.生长激素与胰岛素样生长因子-Ⅰ对马氏珠母贝壳生长形成相关基因表达的影响[J].热带海洋学报，2012，31（2）：96-101.
[9] Liu C，Lovenberg TW. Relaxin-3，INSL5，and their receptors [J]. Results Probl Cell Differ，2008，46：213-237.
[10] Meyts PD，Gauguin L，Svendsen AM，et al. Structural basis of allosteric ligand-receptor interations in the insulin/relaxin peptide family：Implications for other receptor tyrosine kinases and G-protein-coupled receptors [J]. ANN NY Acad，2009，1160：45-53.
[11] 季芳.糖尿病药物治疗的研究进展[J].山西医药杂志，2020，49（19）：2582-2584.
[12] Chan SJ，Cao QP，Steiner DF. Evolution of the insulin superfamily：Cloning of a hybrid insulin/insulin-like growth factor cDNA from amphioxus [J]. PNAS，1990，87（23）：9319-9323.
[13] 陈士光，赖璐华，邓珊珊，等.胰岛素样生长因子-1（IGF-1）与2型糖尿病周围神经病变的关系研究[J].吉林医学，2021，42（3）：539-541.
[14] 梁东春，郭刚，左爱军，等.大鼠胰岛素样生长因子I基因的克隆及表达[J].遗传学报，2002，29（12）：1063-1067.
[15] 张建峰，侯加法，张皎，等.鸡类胰岛素生长因子I的克隆、表达及其表达产物的生物学活性研究[J].中国农业科学，2005（10）：2129-2133.
[16] Smit AB，Spijker S，Minnen JV，et al. Expression and characterization of molluscan insulin-related peptide Ⅷ from the mollusc Lymnaea stagnalis [J]. Neuroscience，1996，70（2）：589-596.
[17] Puig O，Tjian R. Transcriptional feedback control of insulin receptor by dFOXO/FOXO1 [J]. Gene Dev，2005， 19（20）：2435-2446.
[18] Awaji M，Moriyama S. Physiological function of the insulin-related peptides in molluscs [J]. Bull. Fish. Res. Agen.，2008，26（1）：77-82.
[19] Hamano K，Awaji M，Usuki H. cDNA structure of an insulin-related peptide in the Pacific oyster and seasonal changes in the gene expression [J]. J Endocrinol，2005， 187（1）：55-67.
[20] 汲康，孙虎山.长牡蛎消化系统胰岛素样蛋白的免疫组织化学定位[J].鲁东大学学报：自然科学版，2017，33（2）：144-147，158.
[21] 赵子涵，王姿曼，邓岳文，等.大珠母贝胰岛素相关肽受体基因的克隆与表达[J].广东海洋大学学报，2020，40（5）：34-42.
[22] Claeys I，Simonet G，Poels J，et al. Insulin-related peptides and their conserved signal transduction pathway [J]. Peptides，2002，23（4）：807-816.
[23] Brogiolo W，Stocker H，Ikeya T，et al. An evolutionarily conserved function of the Drosophila insulin receptor and insulin-like peptides in growth control [J]. Curr Biol，2001，11（4）：213-221.
[24] Okamoto N. 2016，Molluscan Insulin-Related Peptides [J]. Handbook of Hormones，2016，45：374-376.
[25] 张龙辉，王国栋.无脊椎动物胰岛素样蛋白（Insulin-like/related peptides）研究进展——以昆虫为例[J].生物技术通报，2014（10）：33-42.
[26] Gricourt L，Mathieu M，Kellner K. An insulin-like system involved in the control of Pacific oyster Crassostrea gigas reproduction：hrIGF-1 effect on germinal cell proliferation and maturation associated with expression of an homologous insulin receptor-related receptor [J]. Aquaculture，2006，251（1）：85-98.