Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone in Pichia pastoris
This study was established to test the hypothesis of whether the codon optimization of fish growth hormone gene (FGH) based on P. pastoris preferred codon will improve the quantity of secreted rFGH in culture supernatant that can directly be used as fish feed supplements. The optimized FGH coding se...
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Wiley
2014-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1155/2014/514835 |
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| author | Hussin A. Rothan Teh Ser Huy Zulqarnain Mohamed |
| author_facet | Hussin A. Rothan Teh Ser Huy Zulqarnain Mohamed |
| author_sort | Hussin A. Rothan |
| collection | DOAJ |
| description | This study was established to test the hypothesis of whether the codon optimization of fish growth hormone gene (FGH) based on P. pastoris preferred codon will improve the quantity of secreted rFGH in culture supernatant that can directly be used as fish feed supplements. The optimized FGH coding sequence (oFGH) and native sequence (nFGH) of giant grouper fish (Epinephelus lanceolatus) were cloned into P. pastoris expression vector (pPICZαA) downstream of alcohol oxidase gene (AOX1) for efficient induction of extracellular rFGH by adding 1% of absolute methanol. The results showed that recombinant P. pastoris was able to produce 2.80±0.27 mg of oFGH compared to 1.75±0.25 of nFGH in one litre of culture supernatant. The total body weight of tiger grouper fingerlings fed with oFGH increased significantly at third (P<0.05) and fourth weeks (P<0.01) of four-week experiment period compared to those fed with nFGH. Both oFGH and nFGH significantly enhanced the final biomass and fish survival percentage. In conclusion, codon optimization of FGH fragment was useful to increase rFGH quantity in the culture supernatant of P. pastoris that can be directly used as fish feed supplements. Further studies are still required for large scale production of rFGH and practical application in aquaculture production. |
| format | Article |
| id | doaj-art-65254e33ad6e44e38a0174a4cb07f9c4 |
| institution | Kabale University |
| issn | 2356-6140 1537-744X |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Wiley |
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| series | The Scientific World Journal |
| spelling | doaj-art-65254e33ad6e44e38a0174a4cb07f9c42025-02-03T01:13:13ZengWileyThe Scientific World Journal2356-61401537-744X2014-01-01201410.1155/2014/514835514835Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone in Pichia pastorisHussin A. Rothan0Teh Ser Huy1Zulqarnain Mohamed2Institute of Research Management & Monitoring, Deputy Vice Chancellor (Research & Innovation) Building, University of Malaya, 50603 Kuala Lumpur, MalaysiaGenetics and Molecular Biology Unit, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, MalaysiaGenetics and Molecular Biology Unit, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, MalaysiaThis study was established to test the hypothesis of whether the codon optimization of fish growth hormone gene (FGH) based on P. pastoris preferred codon will improve the quantity of secreted rFGH in culture supernatant that can directly be used as fish feed supplements. The optimized FGH coding sequence (oFGH) and native sequence (nFGH) of giant grouper fish (Epinephelus lanceolatus) were cloned into P. pastoris expression vector (pPICZαA) downstream of alcohol oxidase gene (AOX1) for efficient induction of extracellular rFGH by adding 1% of absolute methanol. The results showed that recombinant P. pastoris was able to produce 2.80±0.27 mg of oFGH compared to 1.75±0.25 of nFGH in one litre of culture supernatant. The total body weight of tiger grouper fingerlings fed with oFGH increased significantly at third (P<0.05) and fourth weeks (P<0.01) of four-week experiment period compared to those fed with nFGH. Both oFGH and nFGH significantly enhanced the final biomass and fish survival percentage. In conclusion, codon optimization of FGH fragment was useful to increase rFGH quantity in the culture supernatant of P. pastoris that can be directly used as fish feed supplements. Further studies are still required for large scale production of rFGH and practical application in aquaculture production.http://dx.doi.org/10.1155/2014/514835 |
| spellingShingle | Hussin A. Rothan Teh Ser Huy Zulqarnain Mohamed Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone in Pichia pastoris The Scientific World Journal |
| title | Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone in Pichia pastoris |
| title_full | Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone in Pichia pastoris |
| title_fullStr | Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone in Pichia pastoris |
| title_full_unstemmed | Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone in Pichia pastoris |
| title_short | Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone in Pichia pastoris |
| title_sort | effect of codon optimisation on the production of recombinant fish growth hormone in pichia pastoris |
| url | http://dx.doi.org/10.1155/2014/514835 |
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