Please wait a minute...

中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2011, Vol. 31 Issue (7): 8-13    
研究报告     
慢病毒载体介导GHRH基因在小鼠肌肉组织表达及对动物生长的影响
戴建威1,3, 章倩倩4, 刘松财3, 张明军3, 张永亮2
1. 广州医学院 广州 510182;
2. 华南农业大学动物科学学院 广州 510642;
3. 吉林大学畜牧兽医学院 长春 130062;
4. 中山大学生物工程中心 广州 510275
Lentiviral Vector Mediated GHRH Gene Expression in Mouse Muscle and the Effect on Growth of Animals
DAI Jian-wei1,3, ZHANG Qian-qian4, LIU Song-cai3, ZHANG Ming-jun3, ZHANG Yong-liang2
1. Department of Biochemistry, Guangzhou Medical College, Guangzhou 510182, China;
2. College of Animal Science, South China Agricultural University, Guangzhou 510642, China;
3. Department of Biochemistry, Veterinary College, Jinlin University, Changchun 130062, China;
4. Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory for Biocontrol, Sun Yat-Sen University, Guangzhou 510275, China
 全文: PDF(738 KB)   HTML
摘要:

人与动物体内生长激素受生长激素释放激素(Growth Hormone Releasing Hormone, GHRH)与生长激素抑制激素(Somatostatin,SST)两种因子共同调节,在体内表达外源GHRH,可以提高体内GH基础水平,进而达到促进体内GH释放,加速动物生长的效果。对慢病毒载体系统加以改造,使之成为CMV与SP双启动子共同启动的基因转移载体,并将GHRH连接双启动子载体后,构建表达载体后与包装质粒共转染293T细胞,包装获得LV-SP-GHRH假病毒颗粒。收集病毒颗粒后注射小鼠肌肉组织,观察其对小鼠生长的影响,同时检测病毒结构基因对受感染细胞基因组整合状况,评估分析假病毒安全性。结果发现,注射病毒颗粒28d时小鼠日增重量与对照组相比,提高153.85%(P<0.01);而且受感染细胞基因组中无病毒结构基因整合,提示表达GHRH基因的慢病毒可以显著且安全地促进动物生长。

关键词: 生长激素生长激素释放激素双启动子慢病毒    
Abstract:

Overexpression of Growth hormone-releasing hormone (GHRH) can control the synthesis and secretion of the growth hormone (GH) from the hypophysis as well as regulate the GH concentrations in animals and humans. The lentiviral vector system were modified to be a CMV-SP two promoter vector. GHRH cDNAs were cloned into the vector and transfected 293T cells with other three plasmids to package LV-SP-GHRH pseudovirion. Muscle tissue of mice were injected with the virus particles and the animal weight growth were observed. At the same time viral safety was assessed by detection of whether the virus structural genes integrated into the genome of infected cells. The results showed that the body weight gain per day increased 153.85% compared to the control group at 28th day postinjection, and the genome of infected cells without virus structural gene integration. All the results indicated that lentiviral vector mediated GHRH gene can promote animal growth significantly and safely.

Key words: Growth hormone    Growth hormone releasing hormone    Two promoter    Lentivirus
收稿日期: 2011-01-05 出版日期: 2011-07-25
ZTFLH:  S811.2  
服务  
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章  
戴建威
章倩倩
刘松财
张明军
张永亮

引用本文:

戴建威, 章倩倩, 刘松财, 张明军, 张永亮. 慢病毒载体介导GHRH基因在小鼠肌肉组织表达及对动物生长的影响[J]. 中国生物工程杂志, 2011, 31(7): 8-13.

DAI Jian-wei, ZHANG Qian-qian, LIU Song-cai, ZHANG Ming-jun, ZHANG Yong-liang. Lentiviral Vector Mediated GHRH Gene Expression in Mouse Muscle and the Effect on Growth of Animals. China Biotechnology, 2011, 31(7): 8-13.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I7/8


[1] 张永亮,刘松财,欧阳红生,等.生长激素释放因子(GRF)在动物肌肉组织中的表达.中国兽医学报,2000,20(3):239-241. Zhang Y L, Liu S C, Ouyang H S, et al.Chinese Journal of Veterinary Science, 2000,20(3):239-241.

[2] Zhang Y L, Ren X H, Liu S C, et al. Overexpression of GRF encapsulated in PLGA microspheres in animal skeletal muscle induces body weight gain. Anim Biotechnol, 2007; 18(3): 163-175.

[3] 任晓慧,张永亮,刘松财,等.微球包裹pcDNA3-GHRH(1-32)在小鼠肌肉组织的表达及对生长的影响.生物工程学报,2004,20(4):615-618. Ren X H, Zhang Y L, Liu S C, et al.Chinese Journal of Biotechnology, 2004,20(4):615-618.

[4] 戴建威,刘松财,任晓慧,等.生长抑素与乙肝表面抗原真核表达载体构建及其对Vc獭兔生长的影响.中国兽医学报,2006,26(1):1-4. Dai J W, Liu S C, Ren X H, et al.Chinese Journal of Veterinary Science, 2006,26(1): 1-4.

[5] Dai J W,Liu S C,Zhang Y L. Simultaneous expression of Growth Hormone Releasing Hormone (GHRH) and Hepatitis B Surface Antigen/Somatostatin (HBsAg/SS) fusion genes in a construct in the skeletal muscle enhances rabbit weight gain. Animal Biotechnology, 2008, 19(2): 104-116.

[6] 周芬,程时远.基因治疗中的非病毒载体.化学通报, 2005, 68 (6):1-7. Zhou F, Cheng S Y. Chemistry, 2005,68 (6): 1-7.

[7] Naldini L,Blomer U,Gallay P,et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector.Science,1996,272(5259):263-267.

[8] Blomer U,Naldini L,Kafri T,et al.Highly efficient and sustained gene transfer in adult neurons with a lentivirus vector.J Virol,1997,71(9):6641-6649.

[9] Kafri T,Blomer U,Peterson D A,et al.Sustained expression of genes delivered directly into liver and muscle by lentiviral vectors.Nat Genet,1997,17(3):314-317.

[10] 章倩倩,戴建威,张永亮,等.CMV与SP双启动子增强外源基因在小鼠骨骼肌中的表达效率. 中国生物化学与分子生物学报, 2007, 23(7): 554-559. Zhang Q Q, Dai J W, Zhang Y L. et al.Chinese Journal of Biochemistry and Molecular Biology, 2007, 23(7): 554-559.

[11] Sinn P L,Sauter S L,McCray P B Jr.Gene therapy progress and prospects:development of improved lentiviral and retroviral vectors--design,biosafety,and production.Gene Ther,2005,12(14):1089-1098.

[12] Beaudet M J,Rueda N, Kobinger G P, et al. Construction of a ganciclovir-sensitive lentiviral vector to assess the influence of angiopoietin-3 and soluble Tie2 on glioma growth. J Neurooncol, 2010,99(1): 1-11.

[13] Breckpot K, Escors D, Arce F, et al. HIV-1 lentiviral vector immunogenicity is mediated by Toll-like receptor 3 (TLR3) and TLR7. J Virol, 2010, 84(11): 5627-5636.

[14] Draghia-Akli R, Li X G, Schwatz R J.Enhanced growth by ectopic expression of growth Hormone releasing hormone using an injectable myogenic vector.Nature Biotechnology,1997,15:1285.

[15] Draghia-Akli R, Ellis K M.High-efficiency growth hormone releasing hormone plasmid vector administration into skeletal muscle mediated by electroporation in pigs. The FASEB Journal,2003,17(3):526-528.

[1] 赵晓煜,徐祺玲,赵晓东,安云飞. 基因治疗慢病毒载体的转导增强策略*[J]. 中国生物工程杂志, 2021, 41(8): 52-58.
[2] 钱昱,丁晓雨,刘志强,袁增强. 基因修饰人多能干细胞的高效单克隆建系方法[J]. 中国生物工程杂志, 2021, 41(8): 33-41.
[3] 黄胜, 严启滔, 熊仕琳, 彭弈骐, 赵蕊. 基于CRISPR/Cas9-SAM系统CHD5基因过表达慢病毒载体的构建及对膀胱癌T24细胞增殖,迁移和侵袭能力的影响 *[J]. 中国生物工程杂志, 2020, 40(3): 1-8.
[4] 菅璐,黄映辉,梁天亚,王利敏,马洪涛,张婷,李丹阳,王明连. 利用CRISPR/Cas9技术建立敲除JAK2基因K562细胞系 *[J]. 中国生物工程杂志, 2019, 39(7): 39-47.
[5] 马占兵,党洁,杨继辉,霍正浩,徐广贤. 基于慢病毒系统的双荧光标记多功能自噬流监测系统建立与应用 *[J]. 中国生物工程杂志, 2019, 39(5): 88-95.
[6] 韩亚丽,杨冠恒,陈雁雯,龚秀丽,张敬之. 表达β-珠蛋白基因的安全性慢病毒载体的优化 *[J]. 中国生物工程杂志, 2018, 38(7): 50-57.
[7] 王丽燕, 王煜, 吴坚平, 徐刚, 杨立荣. 腈水合酶NHaseK在大肠杆菌中的功能表达[J]. 中国生物工程杂志, 2016, 36(12): 42-48.
[8] 代玉环, 徐尧, 罗颖, 代洋, 石伟林, 徐瑶. Myocardin调控心肌H9C2细胞Ca2+通道机制研究[J]. 中国生物工程杂志, 2016, 36(11): 1-6.
[9] 高越, 檀硕, 任兆瑞, 张敬之. 原位染色检测慢病毒载体转录通读方法的建立[J]. 中国生物工程杂志, 2015, 35(5): 55-60.
[10] 刘雪杰, 林巍然, 唐立春, 孙薇, 魏汉东, 姜颖. 慢病毒载体介导RAB27A基因过表达对人HepG2肝癌细胞增殖的影响[J]. 中国生物工程杂志, 2014, 34(9): 16-23.
[11] 李玉强, 朱志图, 王巍, 李谌, 徐娜, 王钰, 李男, 孙宏治. RNA干扰NUP88基因对人乳腺癌MCF-7细胞生长及侵袭力的影响[J]. 中国生物工程杂志, 2014, 34(9): 31-39.
[12] 秦瑶, 赵鸿彦, 张文航, 王冬梅. 线粒体转录因子A敲低转基因小鼠的研制[J]. 中国生物工程杂志, 2014, 34(7): 44-48.
[13] 付辉, 李菲菲, 马琼, 付怀秀, 崔玉芳, 毛建平. 逆转录法筛选mRNA靶点设计核酶对GPA的表达干预实验研究[J]. 中国生物工程杂志, 2014, 34(3): 84-90.
[14] 王鑫, 陈玲, 孙飞, 陆航. RNAi沉默CXCR7对人结肠癌细胞SW620特异性靶向抑制的实验研究[J]. 中国生物工程杂志, 2014, 34(2): 14-20.
[15] 闫亚彬, 蔡勤, 蔡琳琳, 龚秀丽, 朱怡文, 管翌华, 黄英. 比较牛催乳素与生长激素对外源基因表达的影响[J]. 中国生物工程杂志, 2013, 33(8): 91-97.