| 1. | We have identified in s . cerevisiae distinct ubiquitin - ligase complexes that define different erad pathways
我们在酵母体内发现了一种参与不同erad途径的特定的泛素连接酶复合体。 |
| 2. | Ligase an enzyme that catalyzes the bond formation between two substrates at the expense of the breakdown of atp or some other nucleotide triphosphate
连接酶：可将两个底物结合在一起的酶，此过程需要atp或其他核苷三磷酸供能。 |
| 3. | Then cdnas and puc18 vectors were linked by t4 dna ligase and transformed into e . coli strain dh5 - alpha to generate cdna library that size is 4 . 9 l06 recombinants
将cdna与载体连接，并导入dh5感受态细胞中，构建成cdna文库。 |
| 4. | As bases are added by polymerase to the starting point of a new complementary strand , known as a primer , or recognized by ligase as a match , the template ' s sequence is revealed
当聚合酶将一个核苷酸加在新互补链的起始引子之后，或接合酶认定某段核苷酸链与原始模版配对，就可利用这些反应来得出原始模版的序列。 |
| 5. | 4 . the construction of middle - clone vector and expression vector the puc - cp and pgem - 7z plasmid were digested by kpnl and bamhi , and collected the digested cpti fragment and the pgem - 7z , then ligated by t4 dna ligase and formed the pgem - cp
中间载体及表达载体的构建将puc - cp质粒和pgem ? 7z质粒，用kpni和bamhi酶切，分别回收cpti片断和酶切后的载体片段，用t _ 4连接酶连接构建成中间载体pgem - cp 。 |
| 6. | Sub - - clone of s , . / hbsag fusion gene : pbuescripts , . / hbsag and ppiczaa were digested separately by xhoi and xbai enzyme , and were linked under t4 dna ligase , ppiczaa s , , / hbsag was constructed and transformed to e . coli
Hbsag质粒与ppiczaa载体分别经xhol和xbaln切，再在t4dna连接酶作用下进行连接，获得工程菌表达型ppiczaas ； hbsag质粒，转化大肠杆菌t0p10细胞，经xhol和xbal与sacll和xbal酶切电泳，证实s ； 。 |
| 7. | Cut off beta fragment from plasmid prok . ii with hindlll and ecor i as insert , and cut pa into linear plasmid as vector fragment . link the insert and vector fragment together with t4 ligase , and the new vector with gene beta and gus was constructed
用hind和ecor双酶切prok质粒，获得beta基因片段作为插入片段，用hind和ecor双酶切a质粒作为载体片段，将插入片段与载体片段相连，即构建成含有beta和gus的双基因载体。 |
| 8. | At first . then eight a - amylase gene fragments were cloned with the genomic dnas as templates by routine pcr . following that , these gene fragments and plasmid vectors , pbluescript ii ks + and puc18 , were cut by bamh i and kpn i . the prepared insert dna and vector dna were linked by t4 dna ligase
利用vectornti6 . 0软件，对所克隆的序列用相邻接点法( neighborjoining州j ） method ）进行多序列比对，分析其同源性，并构建基因进化树。 |
| 9. | The recombinant plasmid puge dna and transfer vector pfastbacl dna were treated again in the same enzyme , were linked by means of t4 dna ligase and transformed into e . coli jm109 permissive cells , yielding recombinant transfer vector plasmid pfastbac - ge dna and were transformed into dhlobac containing vector bacmid
将重组质粒pugedna与转移载体pfastbacldna用bamhi和ecori双酶切处理， t _ 4dna连接酶连接，用连接产物转化大肠杆菌jm109感受态细胞，得到重组转移载体质粒pfastbac - gedna 。 |
| 10. | This study demonstrated that the arabidopsis f - box protein coil associated with atcul1 , atrbxl and skpl - like proteins askl and ask2 to assemble scfcoil ubiquitin ligase complexes . also , we found that the atcull component of scfcoil complexes contained two species including atcull and modified atcull . ( 2 ) we found that coil assembled to two separate scfcoil complexes with either askl or ask2 through immunoprecipitation analysis with plant expressing myc - tagged version of ask2
用表达融合蛋白myc - ask2的拟南芥为材料，以- myc抗体进行免疫共沉淀分析发现， myc - ask2蛋白可以与coi1蛋白一起免疫共沉淀，但是不能与ask1蛋白免疫共沉淀，表明coi1蛋白与ask2蛋白，但是不能同时与ask1结合形成scf ~ ( coi1 )复合体。 |
