| Autor: |
HAN Tian-tian, YIN Jie, ZENG Jing, JIN Ying, LI Yan, PAN Ling-ya |
| Jazyk: |
čínština |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
Jichu yixue yu linchuang, Vol 42, Iss 6, Pp 896-901 (2022) |
| Druh dokumentu: |
article |
| ISSN: |
1001-6325 |
| DOI: |
10.16352/j.issn.1001-6325.2022.06.003 |
| Popis: |
Objective To establish a stable-expressing human cell cycle regulatory protein 50A (CDC50A) SKOV3 cell strain expressing vector containing CDC50A by molecular cloning. Methods CDC50A was introduced into the expression vector pLVX-IRES-GFP by molecular cloning method. The pLVX-CDC50A-GFP expression plasmid was packaged by lentivirus and stably transfected into SKOV3. SKOV3 cell strain with high-expressing CDC50A was established. The expression of Flag protein was analyzed by flow cytometry, Western blot and immunofluorescence. Results The construction of pLVX-CDC50A-GFP expressing vector was confirmed by gel electrophoresis and endonuclease digestion. The efficiency of infection was observed qualitatively under fluorescence microscope and detected quantitatively by flow cytometry as 86.5%. Flag-tag protein was detected by Western blot to present only in the positive group. Conclusions The SKOV3 cell strain was established by infecting with lentivirus of pLVX-CDC50A-GFP, laying a foundation for the mechanism study of CDC50A in ovarian cancer. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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