Abstract
METHOD: Using spinach to extract total RNA, the spinach GO cDNA was amplified by RT-PCR. This cDNA was inserted into cloning vector pMD-T and sequenced. The cDNA was then cloned into pPIC3.5K (P. Pastoris expression vector) by SnaBI/NotI double digestion. RESULT: Screening by PCR, we got the recombinant plasmid pPIC3.5K-GO which was confirmed by SnaBI/NotI restriction enzyme analysis. Sequencing showed that this cDNA was spinach spinach glycolate oxidase cDNA. The SnaBI/NotI double digestion of pPIC3.5K-GO ensured that the construction is correct. CONCLUSION: Obtained spinach glycolate oxidase cDNA and its recombinant plasmid pPIC3.5K-GO which will be useful for further expression study.
| Original language | English |
|---|---|
| Pages (from-to) | 81-84 |
| Number of pages | 4 |
| Journal | Journal of China Pharmaceutical University |
| Volume | 34 |
| Issue number | 1 |
| State | Published - Feb 2003 |
Keywords
- Cloning
- Spinach glycolate oxidase
- Yeast expression vector