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Characterization of transgene integration pattern in F4 hGH-transgenic common carp (Cyprinus carpio L.)
Wu, B; Sun, YH; Wang, YW; Wang, YP; Zhu, ZY; Zhu, ZY, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Peoples R China
2005-06-01
Source PublicationCELL RESEARCH
ISSN1001-0602
Volume15Issue:6Pages:447-454
AbstractThe integration pattern and adjacent host sequences of the inserted pMThGH-transgene in the F4 hGH-transgenic common carp were extensively studied. Here we show that each F4 transgenic fish contained about 200 copies of the pMThGH-transgene and the transgenes were integrated into the host genome generally with concatemers in a head-to-tail arrangement at 4-5 insertion sites. By using a method of plasmid rescue, four hundred copies of transgenes from two individuals of F4 transgenic fish, A and B, were recovered and clarified into 6 classes. All classes of recovered transgenes contained either complete or partial pMThGH sequences. The class I, which comprised 83% and 84.5% respectively of the recovered transgene copies from fish A and B, had maintained the original configuration, indicating that most transgenes were faithfully inherited during the four generations of reproduction. The other five classes were different from the original configuration in both molecular weight and restriction map, indicating that a few transgenes had undergone mutation, rearrangement or deletion during integration and germline transmission. In the five types of aberrant transgenes, three flanking sequences of the host genome were analyzed. These sequences were common carp beta-actin gene, common carp DNA sequences homologous to mouse phosphoglycerate kinase-1 and human epidermal keratin 14, respectively.; The integration pattern and adjacent host sequences of the inserted pMThGH-transgene in the F4 hGH-transgenic common carp were extensively studied. Here we show that each F4 transgenic fish contained about 200 copies of the pMThGH-transgene and the transgenes were integrated into the host genome generally with concatemers in a head-to-tail arrangement at 4-5 insertion sites. By using a method of plasmid rescue, four hundred copies of transgenes from two individuals of F4 transgenic fish, A and B, were recovered and clarified into 6 classes. All classes of recovered transgenes contained either complete or partial pMThGH sequences. The class I, which comprised 83% and 84.5% respectively of the recovered transgene copies from fish A and B, had maintained the original configuration, indicating that most transgenes were faithfully inherited during the four generations of reproduction. The other five classes were different from the original configuration in both molecular weight and restriction map, indicating that a few transgenes had undergone mutation, rearrangement or deletion during integration and germline transmission. In the five types of aberrant transgenes, three flanking sequences of the host genome were analyzed. These sequences were common carp beta-actin gene, common carp DNA sequences homologous to mouse phosphoglycerate kinase-1 and human epidermal keratin 14, respectively.
SubtypeArticle
KeywordTransgenic Common Carp Plasmid Rescue Germline Transmission Integration Pattern
DepartmentChinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Peoples R China; Wuhan Univ, Coll Life Sci, Wuhan 430072, Peoples R China
Subject AreaCell Biology
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaCell Biology
WOS SubjectCell Biology
WOS IDWOS:000230444400005
WOS KeywordPLASMID RESCUE ; DNA ; GENOME ; GENE ; REARRANGEMENTS ; TRANSMISSION ; INHERITANCE ; SEQUENCES ; REVEALS ; SITES
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Cited Times:13[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/9210
Collection期刊论文
Corresponding AuthorZhu, ZY, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Peoples R China
Affiliation1.Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Peoples R China
2.Wuhan Univ, Coll Life Sci, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Wu, B,Sun, YH,Wang, YW,et al. Characterization of transgene integration pattern in F4 hGH-transgenic common carp (Cyprinus carpio L.)[J]. CELL RESEARCH,2005,15(6):447-454.
APA Wu, B,Sun, YH,Wang, YW,Wang, YP,Zhu, ZY,&Zhu, ZY, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Peoples R China.(2005).Characterization of transgene integration pattern in F4 hGH-transgenic common carp (Cyprinus carpio L.).CELL RESEARCH,15(6),447-454.
MLA Wu, B,et al."Characterization of transgene integration pattern in F4 hGH-transgenic common carp (Cyprinus carpio L.)".CELL RESEARCH 15.6(2005):447-454.
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