| 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 Publication | CELL RESEARCH
 |
| ISSN | 1001-0602 |
| Volume | 15Issue:6Pages:447-454 |
| Abstract | 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.; 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. |
| Subtype | Article |
| Keyword | Transgenic Common Carp Plasmid Rescue Germline Transmission Integration Pattern |
| Department | Chinese 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 Area | Cell Biology |
| WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
| Indexed By | SCI |
| Language | 英语 |
| WOS Research Area | Cell Biology |
| WOS Subject | Cell Biology |
| WOS ID | WOS:000230444400005 |
| WOS Keyword | PLASMID RESCUE ; DNA ; GENOME ; GENE ; REARRANGEMENTS ; TRANSMISSION ; INHERITANCE ; SEQUENCES ; REVEALS ; SITES |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.ihb.ac.cn/handle/152342/9210 |
| Collection | 期刊论文 |
| Corresponding Author | Zhu, ZY, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Peoples R China |
| Affiliation | 1.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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