| 1. | The product of pcr named vp6 is approximate 1 . 3kb in length . the vp6 gene was cloned into pmd18 - t vector and sequenced
将其插入克隆载体质粒pmd18 - t的ecorv酶切位点处，构建重组质粒pmd18 - t ? vp6 。 |
| 2. | The amount of the goal protein was evaluated by densitometric scanning . it indicated that the 45ku product of the vp6 gene was 26 . 5 % of total bacterial protein of bl21
表达产物经ni - nta金属螯合层析柱纯化后，得到纯化的his - taggedvp6融合蛋白。 |
| 3. | The results of sequencing showed that jl94 isolate complete gene 6 was 1356bp and had a complete open reading frame which encoded 397 amino acides
对克隆的vp6基因进行序列测定，测序结果显示jl94vp6基因全长1356bp ，含有完整的开放阅读框架，编码397个氨基酸。 |
| 4. | The expressed products were purified by metal ( ni2 + ) chelation affinity chromatography and tested by western - blot . the results showed that his - tagged vp6 fusion proteins have a positive reaction with anti - jl94 serum
采用western - blot对表达产物及纯化产物进行反应原性分析，结果表明所得his - taggedvp6融合蛋白具有较好的抗原反应特异性。 |
| 5. | According to the reported complete vp6 nucleotide sequences of group a prv in genbank , a pair of primers was designed to amplify vp6 gene of jl94 with oligo 4 . 1 software . the segments of complete gene 6 of jl94 were obtained from rt - pcr using dsrna extracted from the virus as template
根据genbank中的多株猪a群轮状病毒vp6蛋白完整基因序列，利用oligo4 . 1设计引物，以jl94rna为模板，通过rt - pcr扩增出长约1 . 3kb的基因片段，命名为vp6 。 |
| 6. | The vp6 gene was subcloned from recombinant plasmid pmd18 - t - vp6 into expression plasmid pet - 30a . the recombinant plasmid pet - 30a - vp6 was transformed into e . coli bl21 ( de3 ) and induced with iptg . not only a fusion protein about 45ku as we expected was found but also several smaller polypeptides were observed
从重组质粒pmd18 - t ? vp6上将vp6基因亚克隆到表达载体质粒pet - 30a ，转化大肠杆菌bl21 ( de _ 3 ) ， iptg诱导以后表达出预计的45ku的融合蛋白，同时还有一些小的蛋白表达出来，光密度扫描对表达产物进行初步定量表明， 45ku融合蛋白占菌体总蛋白的26 . 5 。 |
