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Publication
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| Title of Article |
Comparative of Recombinant Vespa affinis Hyaluronidase Expressed in Different Cloning Vector and their Biological Properties |
| Date of Acceptance |
23 May 2018 |
| Journal |
| Title of Journal |
International Journal of Applied and Physical Sciences (IJAPS) |
| Standard |
OTHER () |
| Institute of Journal |
kkg publications |
| ISBN/ISSN |
2414-8946 |
| Volume |
2018 |
| Issue |
Volume 4, Issue 2 |
| Month |
July |
| Year of Publication |
2018 |
| Page |
38-44 |
| Abstract |
Cloning and expression of recombinant Vespa affinis hyaluronidase (rVesA2) was successfully expressed in Escherichia coli system. The VesA2 gene was cloned into pET-17b and pET-32a cloning vector which had molecular weight 41.71 and 59.0 kDa, respectively. The recombinant plasmid of pET-17b was composed 1.08 kDa his-tag at the N-terminal. The 17.14 kDa of fusion tag; thioredoxin tag, histidine tag, and S-tag, was found in pET-32a. The verified expression conditions of rVesA2 induced under the conditions of 0.1 mM IPTG at 37C for 4 hrs gave the highest
quantity of protein expression. The colony PCR and sequencing analysis were used to verify the rVesA2. The positive clones were detected the hyaluronidase activity by a zymographic gel. Recombinant proteins from both cloning vec-
tors were insoluble. However, the recombinant from pET-32a showed higher solubility than that form pET-17b, after dissolving in 4 M urea solution. This result suggests that the fusion tag increase protein solubility. |
| Keyword |
Hyaluronidase enzyme, hyaluronic acid, banded tiger wasp |
| Author |
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| Reviewing Status |
มีผู้ประเมินอิสระ |
| Status |
ตีพิมพ์แล้ว |
| Level of Publication |
นานาชาติ |
| citation |
false |
| Part of thesis |
true |
| Attach file |
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| Citation |
0
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Journal Publication
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