Expression and analysis of partial functional domains of H11 gene from Haemonchus contortus
Partial fragment of H11 gene containing several functional domains from Haemonchus contortus was cloned and expressed after bioinformatics analysis of H11 with the soft ware. The fragment of H11 gene from 670 bp to 1710 bp was amplified by means of polymerase chain reaction (PCR) with a pair of prim...
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| Format: | Article |
| Language: | English |
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Zhejiang University Press
2008-11-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2008.06.001 |
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| author | ZHANG Hong-li JIANG Xiao-lei ZHOU Qian-jin DU Ai-fang |
| author_facet | ZHANG Hong-li JIANG Xiao-lei ZHOU Qian-jin DU Ai-fang |
| author_sort | ZHANG Hong-li |
| collection | DOAJ |
| description | Partial fragment of H11 gene containing several functional domains from Haemonchus contortus was cloned and expressed after bioinformatics analysis of H11 with the soft ware. The fragment of H11 gene from 670 bp to 1710 bp was amplified by means of polymerase chain reaction (PCR) with a pair of primers which was designed according to the published data of our laboratory on GenBank and was named HPS(H11 partial sequence). Then the HPS gene was ligated to the prokaryotic expression vector pET-28b and tansformated into E. coli BL21 (DE3). The positive clones were verified by sequencing. Sequence analysis shows that the gene obtained shared 100% and 98.8% identity to the published full coding sequence of H11 gene from Haemonchus contortus ZJ strain and the reference strain, respectively. And the positive clone was induced by IPTG. The results indicate that the fusion protein was calculated to be about 37.34 kDa by SDS-PAGE. The expression quantity of the recombinant fusion protein could reach 58.9% after being induced for 6 h. Western-blotting analysis with anti-his antibodies against the fusion protein showed the recombinant protein shared specificity. |
| format | Article |
| id | doaj-art-ab88e12e2107458fa9576552726171a1 |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2008-11-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-ab88e12e2107458fa9576552726171a12025-08-20T03:58:14ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552008-11-013459159610.3785/j.issn.1008-9209.2008.06.00110089209Expression and analysis of partial functional domains of H11 gene from Haemonchus contortusZHANG Hong-liJIANG Xiao-leiZHOU Qian-jinDU Ai-fangPartial fragment of H11 gene containing several functional domains from Haemonchus contortus was cloned and expressed after bioinformatics analysis of H11 with the soft ware. The fragment of H11 gene from 670 bp to 1710 bp was amplified by means of polymerase chain reaction (PCR) with a pair of primers which was designed according to the published data of our laboratory on GenBank and was named HPS(H11 partial sequence). Then the HPS gene was ligated to the prokaryotic expression vector pET-28b and tansformated into E. coli BL21 (DE3). The positive clones were verified by sequencing. Sequence analysis shows that the gene obtained shared 100% and 98.8% identity to the published full coding sequence of H11 gene from Haemonchus contortus ZJ strain and the reference strain, respectively. And the positive clone was induced by IPTG. The results indicate that the fusion protein was calculated to be about 37.34 kDa by SDS-PAGE. The expression quantity of the recombinant fusion protein could reach 58.9% after being induced for 6 h. Western-blotting analysis with anti-his antibodies against the fusion protein showed the recombinant protein shared specificity.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2008.06.001<italic>Haemonchus contortus</italic><italic>H11</italic>expression |
| spellingShingle | ZHANG Hong-li JIANG Xiao-lei ZHOU Qian-jin DU Ai-fang Expression and analysis of partial functional domains of H11 gene from Haemonchus contortus 浙江大学学报. 农业与生命科学版 <italic>Haemonchus contortus</italic> <italic>H11</italic> expression |
| title | Expression and analysis of partial functional domains of H11 gene from Haemonchus contortus |
| title_full | Expression and analysis of partial functional domains of H11 gene from Haemonchus contortus |
| title_fullStr | Expression and analysis of partial functional domains of H11 gene from Haemonchus contortus |
| title_full_unstemmed | Expression and analysis of partial functional domains of H11 gene from Haemonchus contortus |
| title_short | Expression and analysis of partial functional domains of H11 gene from Haemonchus contortus |
| title_sort | expression and analysis of partial functional domains of h11 gene from haemonchus contortus |
| topic | <italic>Haemonchus contortus</italic> <italic>H11</italic> expression |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2008.06.001 |
| work_keys_str_mv | AT zhanghongli expressionandanalysisofpartialfunctionaldomainsofh11genefromhaemonchuscontortus AT jiangxiaolei expressionandanalysisofpartialfunctionaldomainsofh11genefromhaemonchuscontortus AT zhouqianjin expressionandanalysisofpartialfunctionaldomainsofh11genefromhaemonchuscontortus AT duaifang expressionandanalysisofpartialfunctionaldomainsofh11genefromhaemonchuscontortus |