| 摘要: |
| 研究了根癌农杆菌LBA 4404和C58C1生长的培养基类型、菌株生长状态、转化溶液、质粒与感受态细胞共放置时间、质粒加入量、液氮速冻时间及热激条件对质粒ProkⅡ转化的影响。结果表明,以70 mmol/L CaCl2作为转化溶液,YEB培养基制备的、OD600为0.5的LBA 4404感受态细胞与10 ng质粒DNA,于0 ℃共放置30 min,液氮速冻5 min,37 ℃热激5 min时转化率最高,达1.40×105μg-1;以70 mmol/L CaCl2作为转化溶液,YEB培养基制备的、OD600为0.5～0.6的C58C1感受态细胞与20 ng质粒DNA,于0 ℃共放置10 min,液氮速冻1 min,37 ℃热激1 min时转化率最高,达1.33×105 μg-1。 |
| 关键词: 根癌农杆菌 感受态细胞 质粒ProkⅡ 转化率 |
| DOI： |
| 分类号: |
| 基金项目:陕西省科技攻关项目（2003K03-G1-04） |
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| Studies on the transformation of plasmid Prok Ⅱ into Agrobacterium tumefaciens LBA4404 and C58C1 |
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| Abstract: |
| Factors including different mediums,growth status,transformation solutions，mixing time of competent cells and plasmid，amount of DNA，frozen and heat shock were evaluated on transformation of plasmid ProkⅡ.The results suggested that the LBA4404 competent cells can be obtained when A.tumefaciens grew at OD600=0.5 with YEB medium,prepared with 70 mmol/L CaCl2,and LBA4404 competent cells mixed with 10 ng plasmid DNA at 0 ℃ for 30 min,then frozen by liquid N2 for 5 min,followed by heat shock at 37 ℃ for 5 min,the transformation frequency reached 1.40×105 μg-1;C58C1 competent cells can be obtained when A.tumefaciens grew at OD600=0.5-0.6 with YEB medium,prepared with 70 mmol/L CaCl2,and C58C1 competent cells mixed with 20 ng plasmind for 10 min,frozen for 1 min,heat shock at 37 ℃ for 1 min,the transformation frequency reached 1.33×105 μg-1. |
| Key words: Agrobacterium tumefaciens competent cell plasmid ProkⅡ transformation frequency |
