THP基因的重新克隆及草菇表达载体的构建

来源 :微生物学报 | 被引量 : 0次 | 上传用户:as5881348
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
PCR technique was used for amplifying THP gene in an unknown vector with primer AFP1 and AFP2.Then THP gene was ligated to pGEM T-Vector to be the plasmid pGTHP4.The plasmid pCAMBIA1301 was digested with restriction enzyme BstEⅡ and NcoⅠ,and digestion product was separated with 1% of agarose gel,then big fragment containing promoter was isolated and purified with the Agarose Gel DNA Extraction Kit.At the same way,the plasmid pGTHP4 was digested with restriction enzyme BstZⅠ and NcoⅠ,and the small fragment containing THP gene was purified from 1% agarose gel with the Agarose Gel DNA Extraction Kit.The big fragment and the small fragment were ligated at Nco Ⅰ digested cohesive-end.The ligation product was re-ligated to be cyclic plsmid by addition to a specific adapter,resulting in the pCTH823,a expression vectorof V.volvacea. PCR technique was used for amplifying THP gene in an unknown vector with primer AFP1 and AFP2.Then THP gene was ligated to pGEM T-Vector to be the plasmid pGTHP4. Plasmid pCAMBIA1301 was digested with restriction enzyme BstEII and NcoI, and digestion product was separated with 1% of agarose gel, then big fragment containing promoter was isolated and purified with the Agarose Gel DNA Extraction Kit. At the same way, the plasmid pGTHP4 was digested with restriction enzyme BstZI and NcoI, and the small fragment containing THP gene was purified from 1% agarose gel with the Agarose Gel DNA Extraction Kit. The big fragment and the small fragments were ligated at Nco I digested cohesive-end. ligation product was re-ligated to be cyclic plsmid by addition to a specific adapter, resulting in the pCTH823, a expression vector of V. volvacea.
其他文献
目的 调查围手术期乳腺癌患者配偶的生存质量及心理应激水平,并分析其生存质量与心理应激的相关性.方法 采用自制的一般资料问卷、WHO生存质量测定简表(WHOQOL-BREF)和亲属应
目的 探讨康惠尔透明贴治疗丹毒的临床效果.方法 将44例丹毒患者按随机数字表法随机分成观察组(n=22)和对照组(n=22).观察组用康惠尔透明贴,对照组用50%硫酸镁湿敷于患处,比较
目的:探讨舒适护理在蛛网膜下腔出血(SAH)患者中的应用效果.方法:将66例SAH患者随机分为实验组和对照组各33例,对照组给予积极治疗及神经内科常规护理,实验组在此基础上进行
目的 探讨综合护理干预对老年体位性低血压的防控效果.方法 对334例老年男性进行问卷调查,根据是否发生体位性低血压将其分为体位性低血压组和非体位性低血压组,比较两组患者
目的探讨用中西药在体外消除肠道杆菌携带的耐药质粒pRST98的可能性.方法将原存在于伤寒杆菌中的耐药质粒pRST98经接合转移分别导入大肠杆菌和鼠伤寒杆菌,选用双黄连和氧氟沙
在内蒙古半干旱草原区大针茅+羊草退化草场进行了围栏封育、机械重耙、松土、浅耕翻、浅耕翻+补播、施肥等项改良措施,经过五年的试验研究,表明这些改良措施都有不同程度的增
实验探讨了快速检测抗除草剂拿捕净(Sethoxydim)基因流的方法,该方法核心是用双氧水和赤霉素GA_3)打破青狗尾草种子的休眠,让其在含适宜剂量的除草剂基质上发芽,在暗处生长4d
为了充分利用贵州高海拔山区夏秋气候凉爽、蔬菜虫害较少、环境不污染或污染小的优势 ,生产秋淡市场需求量较大的无公害四季豆 ,近年来我们在大方、毕节、龙里等县 (市 )高海
实测 2 39株凹叶厚朴样木的胸径和材积 ,采用遗传算法、三次设计法和改进单纯形法等拟合一元材积方程并与对数线性化最小二乘法进行比较。结果表明 :采用遗传算法、三次设计
在已建立的单跨架空索道多荷重状态下的悬链线算法数学模型的基础上 ,采用 Turbo C编程 ,能进行单跨架空索道在单荷重或多荷重状态下的承载索设计计算 ,对索道侧型进行验算和