详细信息
重组人粒细胞集落刺激因子(rhG-CSF)基因在鱼腥藻中的克隆
Cloning of the Gene Encoding the Recombinant Human Granulocyte Colony-Stimulating Factor in Anabaena
文献类型:期刊文献
中文题名:重组人粒细胞集落刺激因子(rhG-CSF)基因在鱼腥藻中的克隆
英文题名:Cloning of the Gene Encoding the Recombinant Human Granulocyte Colony-Stimulating Factor in Anabaena
作者:李清艳[1,2];施定基[3,4];陈翠丽[5,4];赵兴贵[3];邓元告[3];张越男[3];吕金印[1]
第一作者:李清艳
机构:[1]西北农林科技大学生命科学学院;[2]天津科技大学海洋科学与工程学院,天津300450;[3]天津科技大学海洋科学与工程学院;[4]中国科学院植物研究所,北京100093;[5]北京联合大学应用文理学院
第一机构:西北农林科技大学生命科学学院
年份:2005
卷号:25
期号:7
起止页码:1389-1394
中文期刊名:西北植物学报
外文期刊名:Acta Botanica Boreali-occidentalia Sinica
收录:CSTPCD;;北大核心:【北大核心2004】;CSCD:【CSCD2011_2012】;
基金:天津市科技重点攻关项目资助(043182711)
语种:中文
中文关键词:人粒细胞集落刺激因子;鱼腥藻(Anabaena;sp.)PCC;7120;载体;转基因鱼腥藻;三亲接合转移
外文关键词:human granulocyte colony-stimulating factor; Anabaena sp.PCC 7120;vector;transgenic Anabaena ;triparental conjugative transfer
摘要:为了将rhG-CSF基因在鱼腥藻PCC7120中克隆,用于制备口服制剂,利用DNA重组技术,在不改变阅读框的前提下,将hG-CSF基因进行突变,并插入到pUC-19载体上,构建中间载体pUC-G-CSF;将pUC-G-CSF插入到pRL-489的启动子PpsbA的下游,构建穿梭表达载体pRL-G-CSF;通过三亲接合转移方法,将pRL-G-CSF转入丝状体蓝藻鱼腥藻PCC7120内。本试验得到了有抗生素抗性的鱼腥藻,并用PCR技术检测到rhG-CSF基因在转基因鱼腥藻中存在。
The objective is to clone the gene encoding the recombinant human granulocyte colony-stimulating factor in Anabaena sp.PCC 7120 for preparing oral drug.On the principle of codon degeneracy,the codons of the first four amino acids of G-CSF were altered by redesigning PCR primers and a rhG-CSF (cDNA) mutant was gained.The middle vector pUC-G-CSF was gained by cloning the correct gene into vector pUC-19;And then, the middle vector pUC-G-CSF was inserted in pRL-489,after the promoter PpsbA to construct shuttle expression vector pRL-G-CSF.The vector pRL-G-CSF was introduced into Anabaena sp.PCC 7120 by triparental conjugative transfer.The transformation state was confirmed by PCR amplification.
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