乙二醇对体内外源基因表达的作用研究

doi:10.13560/j.cnki.biotech.bull.1985.2018-0675

生物技术通报 ›› 2019, Vol. 35 ›› Issue (4): 64-68.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0675

乙二醇对体内外源基因表达的作用研究

王棋文¹, 李盼¹, 潘翠云¹, 韩芬霞²

1. 河南师范大学生命科学学院省部共建细胞分化调控国家重点实验室培育基地,新乡 453007;
2. 河南科技学院动物科技学院,新乡 453003

收稿日期:2018-07-25 出版日期:2019-04-26 发布日期:2019-05-05
作者简介:王棋文,男,博士,讲师,研究方向：细胞生物学;E-mail：wangqiwen@htu.edu.cn
基金资助:
河南省高等学校重点科研项目计划（19A180019,15A230001）,河南师范大学国家级科研项目培育基金项目（2016PL21）

Effect of Ethylene Glycol on the Expression of Exogenous Genes in Vivo

WANG Qi-wen¹, LI Pan¹, Pan Cui-yun¹, HAN Fen-xia²

1. State Key Laboratory Cultivation Base for Cell Differentiation Regulation,College of Life Science,Henan Normal University,Xinxiang 453007;
2. College of Animal Science and Veterinary Medicine,Henan Institute of Science and Technology,Xinxiang 453003

Received:2018-07-25 Published:2019-04-26 Online:2019-05-05

摘要/Abstract

摘要： 探索在液压转基因中乙二醇对小鼠肝脏外源基因表达的影响。以小鼠液压转基因的条件从小鼠尾静脉注射含不同浓度乙二醇的 pEGFP-C1 质粒溶液,分别在尾静脉注射后24 h、48 h和72 h 取小鼠肝脏右叶制冷冻切片,在波长488 nm的荧光显微镜下观察,统计绿色荧光蛋白表达率;同时进行苏木素染色,进行小鼠肝脏形态结构观察。结果显示,乙二醇浓度在0.05 mol/L-0.5 mol/L 范围内,转基因后24 h时随着乙二醇浓度升高,绿色荧光蛋白阳性细胞率呈降低趋势;72 h 时绿色荧光蛋白阳性细胞率随着乙二醇浓度升高而升高,且在 72 h乙二醇浓度为 0.5 mol/L 时阳性细胞率较高。乙二醇在液压转基因中能调节外源基因的表达,使表达高峰向后推移。

关键词: 小鼠, 液压转基因, pEGFP-C1, 乙二醇

Abstract: This work aims to explore the effect of ethylene glycol on the expression of exogenous genes in mouse liver in hydrodynamics-based transgene. The pEGFP-C1 plasmid solution containing different concentrations of ethylene glycol was injected into the tail vein of mice by hydrodynamics-based transgene. The frozen slices of the right lobe of the mice liver were taken at 24 h,48 h and 72 h after the tail vein injection,and were observed with a fluorescence microscope at 488 nm excitation wavelength and the green fluorescent protein（GFP）expression was quantified. At the same time,hematoxylin staining was also carried out and the morphology of the mouse liver was observed. The positive cell rate of GFP at 24 h after transgenic tended to decrease with the increase of ethylene glycol concentration when the concentration of ethylene glycol ranged in 0.05-0.5 mol/L;while the positive cell rate of GFP at 72 h increased with the concentration of ethylene glycol,and was quite high at concentration of ethylene glycol of 0.5 mol/L. In conclusion,ethylene glycol may regulate the expression of exogenous genes in hydrodynamics-based transgene,and the expression peaks shift backwards.

Key words: mice, hydrodynamics-based transgene, pEGFP-C1, ethylene glycol

王棋文, 李盼, 潘翠云, 韩芬霞. 乙二醇对体内外源基因表达的作用研究[J]. 生物技术通报, 2019, 35(4): 64-68.

WANG Qi-wen, LI Pan, Pan Cui-yun, HAN Fen-xia. Effect of Ethylene Glycol on the Expression of Exogenous Genes in Vivo[J]. Biotechnology Bulletin, 2019, 35(4): 64-68.

参考文献

[1] Liu F, Song Y, Liu D.Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA[J]. Gene Therapy, 1999, 6(7):1258-1266.
[2] Hubner EK, Lechler C, Rosner TN, et al.Constitutive and inducible systems for genetic in vivo modification of mouse hepatocytes using hydrodynamic tail vein injection[J]. Journal of Visualized Experiments Jov, 2018, (132):e56613.
[3] Yamazaki T, Nagashima M, Ninomiya D, et al.Neutralizing antibodies induced by gene-based hydrodynamic injection have a therapeutic effect in lethal influenza infection[J]. Frontiers in Immunology, 2018, 9:47.
[4] Hamachi K, Hayashi H, Shimamura M, et al.Glycols modulate terminator stem stability and ligand-dependency of a glycine riboswitch[J]. Biosystems, 2013, 113(2):59-65.
[5] Flessau S, Wolter C, Poselt E, et al.Fluorescence spectroscopy of individual semiconductor nanoparticles in different ethylene glycols[J]. Phys Chem Chem Phys, 2014, 16(22):10444-10455.
[6] Adolph EJ, Nelson CE, Werfel TA, et al.Enhanced performance of plasmid DNA polyplexes stabilized by a combination of core hydrophobicity and surface PEGylation[J]. J Mater Chem B, 2014, 2(46):8154-8164.
[7] 郭学强. 急性肝功能衰竭相关基因和蛋白的研究[D]. 新乡:河南师范大学, 2016.
[8] Hibbitt OC, Harbottle RP, Waddington S N, et al.Delivery and long-term expression of a 135 kb LDLR genomic DNA locus in vivo by hydrodynamic tail vein injection[J]. J Gene Med, 2007, 9(6):488-497.
[9] Wesche-Soldato DE, Lomas-Neira J, Perl M, et al.Hydrodynamic delivery of siRNA in a mouse model of sepsis[J]. Methods Mol Biol, 2008, 442:67-73.
[10] Kamimura K, Yokoo T, Abe H, et al.Image-guided hydrodynamic gene delivery:current status and future directions[J]. Pharmaceutics, 2015, 7(3):213-223.
[11] 徐存栓, 邢雪琨, 谢来峰, 等. 液压转基因技术应用于大鼠肝脏转基因实验[J]. 解剖学报, 2009, 40(1):103-107.
[12] 孙树森, 赵志刚. 临床药师与药物中毒:甲醇、乙二醇和异丙醇[J]. 药品评价, 2017, 14(2):14-20.
[13] 朱士胜, 黄伟, 赵鹏, 等. SD大鼠急性乙二醇中毒的法医病理学研究[J]. 重庆医科大学学报, 2014, 38(1):43-46.
[14] 郑丽丽, 王冲, 文曙, 等. 聚乙二醇修饰甲硫氨酸脑啡肽静脉注射时对热板致痛小鼠的镇痛作用[J]. 中华临床医师杂志(OL/J), 2013, 7(7):3040-3042.
[15] Lovric M, Granic P, Cubrilo-Turek M, et al.Ethylene glycol poisoning[J]. Forensic Science International, 2007, 170(2-3):213-215.
[16] 胡芳芳, 何钉铃, 宁章勇, 等. 低剂量乙二醇对犬器官的损伤[J]. 中国兽医学报, 2015, 35(12):2027-2031.

乙二醇对体内外源基因表达的作用研究

Effect of Ethylene Glycol on the Expression of Exogenous Genes in Vivo

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	潘国强, 吴思源, 刘璐, 郭惠明, 程红梅, 苏晓峰. 大丽轮枝菌（Verticillim dahliae）突变体库的构建与分析[J]. 生物技术通报, 2023, 39(5): 112-119.
[2]	丁亚群, 丁宁, 谢深民, 黄梦娜, 张昱, 张勤, 姜力. Vps28基因敲除小鼠模型的构建及其对泌乳和免疫性状影响的研究[J]. 生物技术通报, 2022, 38(3): 164-172.
[3]	谢绍怡, 蒋璐蔓, 杨晓峰, 张仙玉, 吴珍芳, 李紫聪. 切割小鼠X染色体的CRISPR/Cas9系统表达载体的构建及验证[J]. 生物技术通报, 2021, 37(5): 67-75.
[4]	张静, 熊燕, 华永琳, 郭玉, 熊显荣, 字向东, 李键. 小鼠骨骼肌纤维类型定量PCR内参基因的筛选[J]. 生物技术通报, 2021, 37(2): 71-79.
[5]	石利霞, 高松枫, 朱蕾蕾. PET水解酶的研究进展[J]. 生物技术通报, 2020, 36(10): 226-236.
[6]	李闯, 文正, 刘畅, 邬敏辰. 菜豆环氧水解酶归一性水解间硝基环氧苯乙烷的研究[J]. 生物技术通报, 2020, 36(1): 73-80.
[7]	张达秀, 贺双丽, 王倩, 蒲仕明, 吴琼. 青年和老年小鼠棕色脂肪来源间充质干细胞生物学特性比较[J]. 生物技术通报, 2019, 35(2): 137-142.
[8]	耿慧君, 邹伟, 崔惠敬, 李晓宇, 王丽丽, 徐永平. 基于转录组学的金黄色葡萄球菌噬菌体安全性评估[J]. 生物技术通报, 2019, 35(12): 64-75.
[9]	董维鹏,王君实,张少华,燕炯. CRISPR系统及其应用于小鼠的研究进展[J]. 生物技术通报, 2018, 34(5): 57-63.
[10]	王跃强,AvinashBhandoola. 小鼠LSK及DN原代细胞中基因编辑方法的建立[J]. 生物技术通报, 2018, 34(5): 64-70.
[11]	张雪,陈亮亮,戴红霞,张文胜,任文燕. 利用CRISPR/Cas9系统在小鼠胚胎干细胞中进行miR-22的功能研究[J]. 生物技术通报, 2018, 34(5): 71-79.
[12]	袁白银, 刘钟颖. EDU脉冲追踪小鼠心脏第二心场细胞迁移方法的优化[J]. 生物技术通报, 2018, 34(12): 84-89.
[13]	袁白银, 胡继盛, 刘钟颖, 黄霞. 敲除Wdr1抑制小鼠原代血管平滑肌细胞的迁移和增殖[J]. 生物技术通报, 2018, 34(12): 179-185.
[14]	闻崇炜,赵烨清,石莉,欧阳臻. 聚乙二醇沉淀联用双水相萃取法纯化蛋清溶菌酶的研究[J]. 生物技术通报, 2017, 33(5): 89-93.
[15]	孟北乾, 张惠文, 张国俊, 徐明恺, 李旭, 张成刚. 豚鼠和BALB/c小鼠对SEC2超抗原作用敏感性的比较[J]. 生物技术通报, 2015, 31(9): 224-231.