Development of a positive-negative selection procedure for gene targeting in fish cells

文献类型： 外文期刊

第一作者： Chen, SL

作者： Chen, SL;Hong, YH;Schartl, M

作者机构：

关键词： Fish;Gene targeting;Homologous recombination vector;Medaka oryzias-latipes;Embryonic stem-cells;Germ-line chimeras;Homologous recombination;Melanoma;Xiphophorus;Establishment;Zebrafish;Cultures;Expression

期刊名称：AQUACULTURE （ 影响因子：4.242； 五年影响因子：4.723 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： The gene targeting technique is a powerful tool for analyzing functions of cloned genes and for generating transgenic animals with site-directed integration of foreign genes. In order to develop this technique in fish, positive-negative selection (PNS) and homologous recombination vectors were constructed, and their expression was examined in fish cells. A vector (pNK) for PNS consists of the neomycin resistance gene (neo) as a positive selectable marker gene and the herpes simplex virus (HSV) thymidine kinase (tk) gene as a negative selectable marker gene. Positive selection with geneticin (G418) of epithelioma papulosum of carp (EPC) cells transfected with linearized pNK vector yielded 350 colonies, while double selection of transfected EPC cells with G418 and gancyclovir (Gc) resulted in nearly complete cell death, demonstrating that the PNS procedure is effective in fish cells. Homologous recombination vectors consist of the Xiphophorus melanoma receptor kinase (X mrk(Y)) gene as homologous sequence in addition to the neo and tk genes. Conditions for homologous recombination vector transfection and drug selection were established. After verification of the feasibility of expression of homologous recombination vectors in EPC cells, the first gene targeting experiments were attempted in the Xiphophorus melanoma cell line, PSM. Positive-negative selection of the targeting vector-transfectants led to a low enrichment in this particular cell line. The reasons for the low enrichment in PSM cells were discussed.

分类号： S9

  • 相关文献

[1]Derivation of a pluripotent embryonic cell line from red sea bream blastulas. Chen, SL,Ye, HQ,Sha, ZX,Hong, Y.

[2]Establishment of a pluripotent embryonic cell line from sea perch (Lateolabrax japonicus) embryos. Cher, SL,Sha, ZX,Ye, HQ.

[3]Inactivated Sendai Virus Suppresses Murine Melanoma Growth by Inducing Host Immune Responses and Down-regulating beta-catenin Expression. Zhang Quan,Zhai Guo Qin,Xue Zheng Feng,Xu Xiang Ming,Yuan Wei Feng,Zhu Hong Fei,Zhu Shan Yuan,Xu Xiang Ming. 2012

[4]Establishment and characterization of a fibroblast cell line derived from Texel sheep. Li, Linfeng F.,Guan, Weijun J.,Li, Han,Zhou, Xueyan Z.,Ma, Yuehui H.,Li, Linfeng F.,Bai, Xiujuan J.. 2009

[5]Establishment and characterization of a fibroblast cell line derived from Mongolian sheep. Liu, Chang-Qing,Guan, Wei-Jun,Ma, Yue-Hui,Liu, Chang-Qing,Guo, Yu. 2011

[6]Large-scale production of enhancer trapping lines for rice functional genomics. Yang, YZ,Peng, H,Huang, HM,Wu, JX,Ha, SR,Huang, DF,Lu, TG.

[7]Induction of somatic embryogenesis and plant regeneration from cotyledon and hypocotyl explants of Eruca sativa Mill.. Zhang, T,Cao, ZY,Wang, XY.

[8]Gene targeting using the Agrobacterium tumefaciens-mediated CRISPR-Cas system in rice. Xu, Rongfang,Li, Hao,Qin, Ruiying,Wang, Lu,Li, Li,Wei, Pengcheng,Yang, Jianbo,Li, Hao,Wei, Pengcheng,Xu, Rongfang,Li, Li. 2014

[9]Gene knockdown by intro-thoracic injection of double-stranded RNA in the brown planthopper, Nilaparvata lugens. Ding, Zhiping,Liu, Zewen,Liu, Shuhua,Yang, Baojun,Zhang, Chengwei.

[10]Recent advances in the development of new transgenic animal technology. Miao, Xiangyang,Miao, Xiangyang.

[11]Generation of a Floxed Allele of the Mouse MicroRNA-200 Clusters. Ren, Hongyan,Bi, Yanzhen,Zheng, Xinmin,Tao, Cong,Li, Kui.

[12]Expression of hGM-CSF in silk glands of transgenic silkworms using gene targeting vector. Xue, Renyu,Cao, Guangli,Gong, Chengliang,Xue, Renyu,Chen, Huimei,Cui, Linlin,Cao, Guangli,Zheng, Xiaojian,Gong, Chengliang,Zhou, Wenlin. 2012

[13]New genetic mechanism, origin and population dynamic of bovine ephemeral fever virus. He, Cheng-Qiang,Liu, Ya-Xin,Ding, Nai-Zheng,Wang, Hong-Mei,Hou, Pei-Li,He, Hong-Bin.

[14]An enhanced vector-free allele exchange (VFAE) mutagenesis protocol for genome editing in a wide range of bacterial species. Gomaa, Ahmed E.,Zhang, Chen,Yang, Zhimin,Shang, Liguo,Jiang, Shijie,Deng, Zhiping,Zhan, Yuhua,Lu, Wei,Lin, Min,Yan, Yongliang. 2017

[15]Variation between Turnip mosaic virus isolates in Zhejiang Province, China and evidence for recombination. Chen, JP,Chen, JP,Adams, MJ. 2002

[16]The Klebsiella pneumoniae nitrogenase Fe protein gene (nifH) functionally substitutes for the chlL gene in Chlamydomonas reinhardtii. Cheng, Q,Day, A,Dowson-Day, M,Shen, GF,Dixon, R. 2005

[17]Construction of a Bacillus thuringiensis engineered strain with high toxicity and broad pesticidal spectrum against coleopteran insects. Liu, Jingjing,Yan, Guixin,Shu, Changlong,Zhao, Can,Song, Fuping,Zhou, Lin,Ma, Junlan,Zhang, Jie,Liu, Chunqin,Huang, Dafang.

[18]Construction of recombinant Marek's disease virus (MDV) lacking the meq oncogene and co-expressing AIV-H9N2 HA and NA genes under control of exogenous promoters. Zhang, Zhenjie,Chen, Wenqing,Ma, Chengtai,Zhao, Peng,Duan, Luntao,Zhang, Fushou,Su, Hongqin,Li, Sifei,Cui, He,Cui, Zhizhong,Sun, Aijun,Li, Yanpeng.

[19]Crossover rate between homologous chromosomes and interference are regulated by the addition of specific unpaired chromosomes in Brassica. Suay, Loreto,Zhang, Deshuang,Eber, Frederique,Jouy, Helene,Lode, Maryse,Huteau, Virginie,Coriton, Olivier,Szadkowski, Emmanuel,Paillard, Sophie,Chevre, Anne-Marie,Zhang, Deshuang,Leflon, Martine,Martin, Olivier C.,Falque, Matthieu,Jenczewski, Eric.

[20]High-Frequency Targeted Mutagenesis in Pseudomonas stutzeri Using a Vector-Free Allele-Exchange Protocol. Gomaa, Ahmed E.,Deng, Zhiping,Yang, Zhimin,Shang, Liguo,Zhan, Yuhua,Lu, Wei,Lin, Min,Yan, Yongliang.

作者其他论文 更多>>

微信小程序