Sulistyarini, Arum (2017) Upaya Peningkatan Solubilitas Protein Rekombinan Ag 38 Kda Mycobacterium Tuberculosis Dengan Metode Heat Shock. Sarjana thesis, Universitas Brawijaya.
Abstract
Antigen 38 (Ag38) merupakan agen virulensi yang dipercaya paling poten menyebabkan dan meningkatkan keparahan tuberkulosis (TB). Gen pab pengkode Ag38 telah diisolasi dan diekspresikan dalam E.coli. Namun, saat protein rekombinan ini diekspresikan, banyak terbentuk protein dalam bentuk yang tidak larut, yang disebabkan terbentuknya badan inklusi (inclusion bodies). Oleh karena itu diperlukan suatu metode yang dapat meningkatkan kelarutan protein sehingga didapatkan protein yang lebih fungsional. Salah satu metode yang dapat meningkatkan kelarutan protein adalah metode heat shock. Penelitian ini bertujuan untuk mengetahui apakah metode heat shock dapat meningkatkan solubilitas protein antigen 38 kDa M. tuberculosis. Metode heat shock dilakukan dengan memapar sel protein terhadap stress panas (heat stress) yang akan memicu ekspresi dari protein heat shock, yang berperan dalam membantu proses refolding, membongkar agregat protein, atau mencegah terbentuknya agregasi protein. Profil protein dengan metode heat shock yang ditunjukkan dari hasil elektroforesis SDS PAGE menunjukkan banyak protein yang berada di pellet, sedangkan yang berada di supernatant hanya sedikit. Artinya bahwa sedikit protein rekombinan yang meningkat kelarutannya. Untuk mendapat protein rekombinan Ag38 kDa murni, dilakukan purifikasi dengan Protino Ni-TED. Kesimpulan dari penelitian ini menunjukkan bahwa metode heat shock tidak bisa membantu meningkatkan kelarutan protein rekombinan Ag38 kDa Mycobacterium tuberculosis.
English Abstract
Antigen 38 (Ag38) the most potent trustworthy virulence agent that causes and increases the severity of tuberculosis (TB). The Ag38 coding genes have been isolated and expressed in E.coli. However, when this recombinant protein is expressed, many proteins form in an insoluble form, due to the formation of inclusion bodies. Therefore, we need a method that can increase the solubility of proteins to obtain a more functional protein. One method that can be applied in improving protein solubility is the heat shock method. Aims of this study is to determine whether heat shock method can increase solubility of 38 kDa M. tuberculosis antigen protein. This method is carried out by exposing the protein cell to heat stress which will trigger the expression of the heat shock protein, which plays a role in helping the refolding process, disassembling protein aggregates, or preventing the formation of protein aggregation. The profile protein with heat shock method shown from electrophoresis result of SDS PAGE shows many proteins residing in pellet, while protein in the supernatant are small. It means that just few amount of recombinant proteins increase their solubility. To obtain a pure Ag38 kDa recombinant protein, purification was performed with the Protino Ni-TED. The conclusions of this study indicate that heat shock method cannot help to increase the solubility of the recombinant protein Ag38 kDa Mycobacterium tuberculosis.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | SKR/FK/2017/466/051800275 |
| Uncontrolled Keywords: | heat shock, Mycobacterium tuberculosis, protein Ag38 kDa, tuberculosis, heat shock, Mycobacterium tuberculosis, Ag38 kDa protein, tuberculosis |
| Subjects: | 600 Technology (Applied sciences) > 616 Diseases > 616.9 Other disease > 616.99 Tumors and miscellaneous communicable diseases > 616.995 Tuberculosis |
| Divisions: | Fakultas Kedokteran > Pendidikan Dokter |
| Depositing User: | Nur Cholis |
| Date Deposited: | 15 Jan 2018 07:15 |
| Last Modified: | 24 Oct 2021 03:40 |
| URI: | http://repository.ub.ac.id/id/eprint/8052 |
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