| 摘要: |
| 目的: 获得HDAC1 的cDNA及构建酵母双杂合系统中的靶基因。方法: 利用RT-PCR方法，从人Jurkat 细胞中扩增出一约1.45kb的DNA片段，作全自动测序,重组入pLexA载体，构建成pLexA-HDAC1, 并用醋酸锂法转化酵母菌EGY48, 在选择性培养基上观察pLexA-HDAC1 在EGY48中的表达情况。结果: 获得的1.45kb的DNA编码的氨基酸序列与文献报道的HDAC1一致，转化的酵母菌在选择性培养基SD/Gal/Raf/-Ura/-His/上培养3d后, 长出约1mm大小的白色菌落。结论：获得了HDAC1的cDNA，pLexA-HDAC1对EGY48没有毒性作用，也没有自身激活LacZ功能，可作为酵母双杂合系统中的靶基因。 |
| 关键词: 组蛋白脱乙酰化酶 酵母双杂交系统 |
| DOI： |
|
| 基金项目: |
|
| Cloning of histone deacetylase 1 cDNA and construction of its yeast expression plasmid |
| 傅继梁,张晓琴,陈坚 |
| () |
| Abstract: |
| Objective: To obtain the histone deacetylase 1(HDAC1) cDNA and construct the target gene of yeast two-hybrid system. Methods: About 1.45kb DNA fragment was amplified from the human Jurkat cell by RT-PCR. After being automaticly sequenced, the fragment was ligased with the vector pLexA to construct pLexA-HDAC1. The recombinant plasmid was transformed into yeast EGY48,and the expression of pLexA-HDAC1 in the yeast was observed. Results: The amino acid sequence encoded by RT-PCR product was the same as reported HDAC1,and about 1 mm white yeast clone grew in the selective medium after 3d. Conclusion:The HDAC1 cDNA has been obtained. pLexA-HDAC1 has nontoxic to yeast, which can serve as a target gene of yeast two-hybrid system. |
| Key words: histone deacetylase yeast two-hybrid system |
