“全鱼”转生长激素基因黄颗鱼首建者的建立*

南京大学学报(自然科学版) ›› 2013, Vol. 49 ›› Issue (1): 123–131.

“全鱼”转生长激素基因黄颗鱼首建者的建立*

葛家春1·2，宋伟3董张及3，许志强1,2，鲍洁3，周国勤4，潘建林2，杨家新1，赵庆顺3**

出版日期:2015-09-17 发布日期:2015-09-17
作者简介: (1.南京师范大学生命科学学院，南京，210046;2.江苏省淡水水产研究所，南京，210017;
3.南京大学模式动物研究所，南京，210061;.南京市水产科学研究所，南京，210036)
基金资助:
江苏省自然科学基金(BK2008484)，江苏省科技支撑计划项日(BE2010376)

Generation of "all fish" growth hormone gene transgenic yellow catfish founders

Ge Jia-Chun1’2，Song Wei3，Dong Zhang一Ji3，X u Zhi一Qiang1.2，Bao Jie3,
Zhou Guo一Qin4，Pan Jian一Lin2,Yang Jia一xin1，Zhao Qing-Shun3

Online:2015-09-17 Published:2015-09-17
About author: (1 .College of Life Scicnces,Nanjing Normal University,Nanjing,210046,China;
2. Freshwater Fisheries Research institute of Jiangsu Province,Nanjing,210017,China;
3. Model Animal Research Center,Nanjing University,Nanjing,210061,China;
4. Nanjing Aquatic Sciences Institute,Nanjing,210036,China)

摘要/Abstract

摘要： 黄颡鱼（Pelteobagrus fulvidraco)是一种重要的淡水名优经济角鱼类，然而较小的体型和较慢
的生长速率极大地降低了它的经济价值.为获得大规格且快速生长的黄颖角，开展了转生长激素基因黄
颖角的研究.运用RT-PCR和RACE-PCR技术，克降了黄颖角生长激素基因的603 bp(碱基对)的编码
序列和485 bp，的3’-UTR(非翻译区)序列.通过重叠PCR和限制性内切酶酶切连接，组建了一个长为
2105 bp的“全鱼”转生长激素基因构件，该构件含1017 bp的黄颡鱼β-肌动蛋白近端启动子、603 bp的
生长激素基因编码序列和485 bp，的3’-UTR序列.采用显微注射将转生长激素基因构件导入黄颖角受
精卵，从301尾由注射肛胎发育而来的黄颖角中共筛选获得40尾转生长激素基因黄颖角首建者.成功
地建立了黄颡鱼基因组改造的技术平台，为黄颡鱼的基因工程育种奠定了基础.

Abstract: Yellow catfish(Pelteobagrus fulivdraco Richardson)is one of the most important freshwater farmed spe-
cies in China. However, its small size and slow growth rate limit its economic value. Urowth hormone is a protein se-
creted by pituitary, It promotes animal growth. Overexpression of growth hormone in fishes results in the growth
hormone gene transgenic fishes with enlarged body size and accelerated growth, In order to increase its body size and
growth rate，we performed transgenic research on yellow catfish. Employing RT-PCR and RACE(rapid amplification
of cDNA ends)-PCR，we cloned 603 bp(base pairs)complete coding sequence and X85 by 3^,-UTR(untranslated re-
gion)of yellow catfish growth hormone gene，respectively. Performing overlapping PLR and molecular cloning with
restriction endonuclcases,we constructed an "all fish" growth hormone gene transgenic construct with 2105 by in
length. The construct comprises 1017 by of yellow catfish份actin proximal promoter, 603 by complete coding se
qucnce and 485 by 3’-UTR of yellow catfish growth hormone gene, respectively. Screening 301 individuals developed
from the yellow catfish fertilized eggs microinjected with the growth hormone gene transgenic construct，we obtained
40 transgenic founders. Our results showed that we have established a yellow catfish transgenic platform that lays
the foundation for our further molecular breeding to create new strains of yellow catfish with large body size and rap-
id growth.

葛家春1·2，宋伟3董张及3，许志强1,2，鲍洁3，周国勤4，潘建林2，杨家新1，赵庆顺3**
. “全鱼”转生长激素基因黄颗鱼首建者的建立*[J]. 南京大学学报(自然科学版), 2013, 49(1): 123–131.

Ge Jia-Chun1’2，Song Wei3，Dong Zhang一Ji3，X u Zhi一Qiang1.2，Bao Jie3,
Zhou Guo一Qin4，Pan Jian一Lin2,Yang Jia一xin1，Zhao Qing-Shun3. Generation of "all fish" growth hormone gene transgenic yellow catfish founders
[J]. Journal of Nanjing University(Natural Sciences), 2013, 49(1): 123–131.

参考文献

[1]Meng Q W,Su J X,Miao X Z. Systematics of fi- shes. Beijing; China Agriculture Press, 1995，
1158.(孟庆闻，苏锦祥，缪学祖.角类分类学.北京:中国农业出版社，1995,1158).
[2]Bjornsson B T. The biology of salmon growth hor- mone;From daylight to dominance. Fish Physiology
and Biochemistry, 1997，17(1):9一24.
[3]Markert J R，Higgs D A，Dye H M, et u1. Influence of bovine growth hormone on growth
rate，appetite, and food conversion of yearling coho salmon(Oncorhynchus kisutch)fed two diets
of different composition. Canadian Journal of Zo- ology-Revue Canadienne De Zoologie, 1977，55 (1):74一83.
[4]Sundstrom L F, Lohmus M, Johnsson J I，et al. Growth hormone transgenic salmon pay for
growth potential with increased predation mortal- ity. Proceedings of the Royal Society B;Biological
Sciences, 2004 , 271 ( Suppl 5 ):S350一S352.
[5]Devlin R. Sequence of sockeye salmon type 1 and 2 growth hormone genes and the relationship of
painbow trout with Atlantic and Pacific salmon. Canadian Journal of Fisheries and Aquatic Sci- ences,l993，50:1738一1748.
[6]Leung T C, Ng T B, Woo N Y. Metabolic effects of bovine growth hormone in the tilapia Oreo-
chromis mossambicus. Comparative Biochemistry and Physiology Part A:Physiology, 1991，99(4): 633一636.
[7]Palmiter R D, Brinster R L, Hammer R E, et al. Dramatic growth of mice that develop from eggs
microinjected with metallothionein-growth hor- mone fusion genes. Nature，1982，300(5893):611 ~615.
[8]Zhu Z，He L,Chen S. Novel gene transfer into the fertilized eggs of gold fish(Carassius aurutus L.1758).Journal of Applied Ichthyology,1985，1(1):31一34.
[9]Zhu Z. Generation of fast growing transgenic fish: Methods and mechanisms. Hew C，Fletcher
G. Transgenic Fish. Singapore; World Scientific Publishing, 1992，92一119.
[10]Du S J，Gong Z Y，Fletcher G L，et al. Growth enhancement in transgenic Atlantic salmon by the
use of an"all fish"chimeric growth hormone gene construct. Biotechnology, 1992，10(2):176一181.
[11]Yaskowiak E S,Shears M A，Agarwal-Mawal A，et al. Characterization and multi-generational sta- bility of the growth hormone transgenc(EO- lalpha)responsible for enhanced growth rates in Atlantic Salmon. Transgenic Research，2006，15(4):465一480.
[12]Devlin R H，Yesaki T Y，Yiagi C A，et al. Ex- traordinary salmon growth. Nature, 1994，371 (6494):209一210.
[13]Martinez R，Estrada M P，Berlanga J，et al. Growth enhancement in transgenic tilapia by ec- topic expression of tilapia growth hormone. Mo- lecular Marine Biology and Biotechnology, 1996，5 (1):62一70.
[14]Rahman M A，Ronyai A，Engidaw B Z, et al.Growth and nutritional trials on transgenic Nile
tilapia containing an exogenous fish growth hor- mone gene. Journal of Fish Biology,2001，59(1): 62一78.
[15]Pitkanen T I，Krasnov A，Teerijoki H，et al. Transfer of growth hormone(GH)transgenes into
Arctic chary.(Sulvelinus alpinus L.)I. Growth response to various GH constructs. Genetic Anal-
ysis:Biomolecular Engineering, 1999，15(3-5):91 ~98.
[16]Dunham R A, Chitmanat C, Nichols A, et al. Predator Avoidance of Transgenic Channel Catfish
vontaining salmonid growth hormone genes. Marine Biotechnology,1999，1(6):545一551,
[17]Devlin R H，Biagi C A，Yesaki T Y,et al. Growth of domesticated transgenic fish. Nature,2001 ,409 (6822):781一782.
[18]Nam Y K，Noh J K，Cho Y S,et al. Dramatically accelerated growth and extraordinary gigantism
of transgcnic mud loach Misgurnus，，mizolepis. Transgenic Research,2001，10(4):353一362.
[19]Deitch E J，Fletcher G L, Petersen L H，et al.Car- diorcspiratory modifications,and limitations,in post-
smolt growth hormone transgenic Atlantic salmon Salmo salar. Journal of Experimental Biology,2006，209(Pt 7):1310一1325.
[20]Fletcher G L，Goddard S V，Shears M, et al. Transgenic salmon;Potential and hurdles. Molec
ular Farming:Proceedings of the OECD Workshop Held in La Grande Motte(France): INRA editions,2000，57一65.
[21]Fletcher G L,Shears M A，Yaskowiak E S,et al. Gene transfer;Potential to enhance the genome of
Atlantic salmon for aquaculture. Australian Journal of Experimental Agriculture, 2004，44 (11):1095一1100.
[22]Ge J，Dong Z, Li J，et al. isolation of yellow catfish beta-actin promoter and generation of
transgenic yellow catfish expressing enhanced yellow fluorescent protein. Transgenic Research, 2012:10一1007.
[23]Sun Y H，Xie C S. Studies on cDNA in the growth hormone of Pelteobagrus fulvidraco. Reservoir
Fisheries, 2006 , 26 ( 3 ) : 10 - 13.(孙玉华，谢从新. 黄颡鱼长激素cDNA序列克降及其序列分析. 水利渔业.2006,26(3):10-13).
[24]Hanky S, Smith T J，Muller F, et al. Isolation and functional analysis of the histone H3 promoter from
atlantic salmon(Sahno salar L.).Molecular Marine Biology and Biotechnology, 1998,7(3):165一172.
[25]Alam M S,Lavender F L,lyengar A,et al. Com- parison of the activity of carp and rat beta-actin
gene regulatory sequences in tilapia and rainbow trout embryos. Molecular Reproduction and De- velopment, 1996,45(2):117一 122.
[26]Udvadia A J,Linney E. Windows into development;Historic, current, and future perspectives on
transgenic zebrafish. Developmental Biology, 2003 ,256(1):1一17.
[27]Dong Z,Ge J,Li K,et al. Heritable targeted inac- tivation of myostatin gene in yellow catfish(Pel-
teobagrus fulvidraco)using engineered zinc finger nucleases. PLoS One,2011，6(12):e28897.

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