摘要
目的：将人特异表达的癌胚抗原（ＣＥＡ）－ｃＤＮＡ克隆到逆转录病毒质粒载体ｐＬＸＳＮ中，构建成真核表达重组质粒ｐＬＸＳＮ－ＣＥＡ，为进一步表达表达人ＣＥＡ的动物肿瘤细胞模型作准备。方法：用内切酶ＥｃｏＲＩ酶切质粒ｐＬＸＳＮ及ｐ９１０２３Ｂ－ＣＥＡ－１７，得到线状载体ｐＬＸＳＮ及外源基因ＣＥＡ－ｃＤＮＡ，然后通过连接酶的连接反应将ＣＥＡ－ｃＤＮＡ插入到ｐＬＸＳＮ的ＥｃｏＲＩ位点，用ＥｃｏＲＩ和ＢａｍＨＩ鉴定。结果：成功地将ＣＥＡ－ｃＤＮＡ克隆到ｐＬＸＳＮ中，通过鉴定筛选获得有意义的正向重组质粒ｐＬＸＳＮ－ＣＥＡ。结论：利用基因克隆技术能将人ＣＥＡ－ｃＤＮＡ定向插入逆转录病毒质粒载体中，构建成真核表达重组质粒ｐＬＸＳＮ－ＣＥＡ，为进一步建立ＣＥＡ阳性动物肿瘤细胞模型及研究ＣＥＡ阳性肿瘤的免疫防治奠定了基础。
Objective: The cDNA of the carcinoembryonic antigen(CEA)
was cloned into pLXSN of the retrovirus plasmid to form eukaryotic expression recombinant
plasmid,pLXSN-CEA. Methods: The pLXSN and p91023B-CEA-17 were digested with EcoRI
separately,and the linear pLXSN and CEA-cDNA fragment were obtained.And then the CEA-cDNA
was inserted into the pLXSN through ligation with ligase. Results: The recombinant plasmid
pLXSN-CEA-cDNA was successfully cloned into the retrovirus vector plasmid pLXSN ,with the
significant recombinant plasmid pLXSN-CEA obtained through screenig. Conclusion: The human
gene CEA-cDNA could efficiently be inserted into retrovirus plasmid to form eukaryotic
expresson recombinat plasmid pLXSN-CEA.This provides the basis for constructing animal CEA
positive rumor cell model and sudying the prevention and therapy of CEA positive tumors.
出处
《郧阳医学院学报》
1999年第2期63-65,共3页
Journal of Yunyang Medical College
关键词
癌胚抗原
克隆
质粒
重组质粒
CDNA
CEA-RV
Carcinoembryonic Antigen
Retrovirus,MLV-Related
Genetic
Vectors
Genes
cloning,Molecular
Plasmids
