Constructing human colorectal cancer cell line with stably down - regulated grp94 ( 1 ) the plasmid prc / rsv - ribol that contains specific grp94 - targeting ribozyme and the control plasmid prc / rsv were miniprepared , respectively , cleaved by endoenzyme pvuii 稳定下调grp94的人大肠癌细胞克隆株的构建( 1 )分别对含有特异性打靶grp94核酶的质粒prc rsv - ribo1和对照组质粒prc rsv进行小量提取、 pvu酶切鉴定。
2.
This time , using cdna of zmcdc5 as template , we amplify a sequence by means of pcr technology . and then , using restrict endoenzyme and ligase , we conjunct the 0 . 8kb length dna sequence in a expression vector , pet - 30a . after induction , expression and purification , we obtained a 35 . 4kda truncated fusing zmcdc5 protein which contains 267aa ( 647 to 914aa in zmcdc5 ) . with the purified protein , we got its antibody and testified the antibody by means of western blotting and dot blotting 本实验是以zmcdc5的cdna为模板，使用pcr获得基因片段，再通过酶切连接，将得到的0 . 8kb的基因片段构建于pet - 30a表达载体上，经过诱导表达和纯化，获得zmcdc5的融合蛋白，其中包括了zmcdc5925个氨基酸中647 914共267个氨基酸残基
