After digested with ecori and noti , the obtained dna fragment was linked with ppic9k by t4dna ligase and then the recombinant plasmid was transformed into competent dhscc . after positive transformants were sieved out by pcr , digesiting analysis and sequencing were also used to confirm the positive result more 所得的dna片段经ecor和not双酶切后用t _ 4dna连接酶与ppic9k载体进行连接，然后导入大肠杆菌dh5 ，用pcr法筛选阳性转化子，并用双酶切和序列测定方法鉴定重组质粒。
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
A pair of primers were designed and synthesized based on the published ge gene sequence of prv - rice strain for amplifying ge gene of prv min - a , yielding a 1 . 7kb band . the segment was linked to puc19 plasma dna by means of t4 dna ligase , transformed into e . coli jm109 permissive cells , and incubated on lb fray containg amp , x - gal and iptg . small amount of plasma was extracted by base cleavaging for enzyme digest analysis and pcr , resulting in recombinant plasma puge dna containing prv ge 用t _ 4dna连接酶使ge基因与经bamhi 、 kpni同样双酶切的puc19质粒dna连接；用连接产物转化大肠杆菌jml09感受态细胞，置含amp 、 x - gal和iptg的lb平板上培养12 20小时；挑取白色菌落于选择性培养基扩大培养，碱裂解法小量提取质粒dna ，并进行酶切分析鉴定，结果获得整合有prvge基因的重组质粒pugedna ，并与其它prv分离株进行ge基因序列同源性分析。
The scfco , " ubiquitin - ligase complexes are required for jasmonate response in arabidopsis ( l ) arabidopsis coi is required for all the jasmonate - regulated response . it encodes a protein containing leucine - rich repeats and a degenerate f - box motif these structural features are characteristic of f - box proteins that function in ubiquitin ligase complexes for the ubiquitylation of substrate - proteins targeted for degradat1on 以表达融合蛋白flag - coi1的拟南芥为材料，用- flag抗体进行免疫共沉淀分析发现， coi1蛋白可以与ask1 、 ask2 、 atrbx1和atcul1蛋白结合形成scf ~ ( coi1 )复合体，并发现其中的atcul1蛋白包括未修饰和修饰的两种形式。
First , the purified pezzis and pcr product of angiostatin are digested by ecor . i and xba i . after purifying the digested products respectively , we ligate these two kinds of dna by t4 dna ligase and construct the recombinant plasmid pezz18 - as . then transform it to the competent e . coli dh5a 用限制性内切酶ecori与xbai对目的基因as 、表达载体pezz18行双酶切，酶切产物纯化后利用大肠杆菌t _ 4dna连接酶连接构成重组子pezz18 - as ，并转化e . colidh5 ，经氨苄青霉素lb平板初筛后，以菌液pcr和重组子的单、双酶切行进一步鉴定。
