NOVITA NUR NAENI, - (2022) OPTIMASI EKSPRESI PROTEIN NANOBODI KORTISOL PADA BAKTERI Escherichia coli BL21(DE3) DENGAN VARIASI KONSENTRASI ISOPROPYL-β-DTHIOGALACTOPYRANOSIDE (IPTG). Skripsi thesis, Sekolah Tinggi Farmasi Indonesia.
![[img]](http://repository.stfi.ac.id/style/images/fileicons/text.png) |
Text
S_PSSF_A181079_Title.pdf Download (530kB) |
|
Text
S_PSSF_A181079_Chapter1.pdf Restricted to Repository staff only Download (233kB) |
|
Text
S_PSSF_A181079_Chapter2.pdf Restricted to Repository staff only Download (460kB) |
|
Text
S_PSSF_A181079_Chapter3.pdf Restricted to Repository staff only Download (246kB) |
|
Text
S_PSSF_A181079_Chapter4.pdf Restricted to Repository staff only Download (659kB) |
|
Text
S_PSSF_A181079_Chapter5.pdf Restricted to Repository staff only Download (138kB) |
|
Text
S_PSSF_A181079_Appendix.pdf Restricted to Repository staff only Download (569kB) |
Abstract
Nanobodi adalah antibodi yang ditemukan dengan ukuran lebih kecil serta mampu berikatan dengan kortisol, sehingga dapat diproduksi sebagai komponen kit diagnostik untuk pemantauan kadar kortisol dalam tubuh. Dalam produksi protein rekombinan, penentuan konsentrasi Isoprophyl-ß-D-thiogalactoside (IPTG) yang optimal sebagai penginduksi dapat menentukan keberhasilan ekspresi protein rekombinan. Penelitian ini bertujuan untuk mengetahui pengaruh variasi konsentrasi IPTG dan mendapatkan konsentrasi yang optimum dalam produksi protein rekombinan pada bakteri E.coli BL21(DE3). Plasmid pET-28a yang mengandung nanobodi natif dan mutan disisipkan ke dalam bakteri menggunakan metode kejut panas. IPTG sebagai penginduksi ditambahkan dengan konsentrasi 0,4; 0,6; 0,8; dan 1 mM pada suhu kamar selama 18 jam kemudian divalidasi dengan SDS PAGE. Hasil penelitian menunjukkan bahwa variasi konsentrasi IPTG berpengaruh terhadap jumlah sel bakteri dengan nilai tertinggi pada nanobodi natif sebanyak 0,469 mg dan nanobodi mutan sebanyak 0,444 mg. Serta menunjukkan tebal pita protein tertinggi pada nanobodi natif dengan nilai AUC 45.324,541 piksel dan nanobodi mutan dengan nilai AUC 28.195,345 piksel. Konsentrasi protein rekombinan tertinggi baik natif maupun mutan diperoleh dengan penambahan induksi IPTG pada konsentrasi 0,6 mM. --- Nanobodies is antibodies that are found with a smaller size and is able to bind to cortisl, so that they can be produced as a component of a diagnostic kit for monitoring cortisol levels in the body. In the production of recombinant proteins, the determination of the optimal concentration of Isoprophyl-ß-D-thiogalactoside (IPTG) as an inducer can determine the success of recombinant protein expression. This study purpose to determine the influence of variations in IPTG concentration and obtain optimum concentrations in the production of recombinant proteins in E.coli BL21 (DE3) bacteria. PET-28a plasmids containing native and mutant nanobodies were inserted into the bacteria using heat shock method. IPTG as an inducer is added with a concentration of 0.4; 0,6; 0,8; and 1 mM at room temperature for 18 hours then validated with SDS PAGE. The results showed that variations in IPTG concentrations affected the number of bacterial cells with the highest value in native nanobodies as much as 0.469 mg and mutant nanobodies as much as 0.444 mg. As well as showing the highest protein band thickness in the native nanobody with an AUC value of 45,324,541 pixels and the mutant nanobody with an AUC value of 28,195,345 pixels. The highest concentrations of recombinant proteins of natives and mutants were obtained by the addition of IPTG induction at a concentration of 0.6 mM.
| Item Type: | Thesis (Skripsi) |
|---|---|
| Uncontrolled Keywords: | Nanobodi, kortisol, Isoprophyl-ß-D-thiogalactoside (IPTG), E.coli BL21(DE3). --- Nanobody, cortisol, Isoprophyl-ß-D-thiogalactoside (IPTG), E.coli BL21(DE3). |
| Subjects: | Q Science > Q Science (General) |
| Divisions: | Program Studi S1 Farmasi |
| Depositing User: | pustakawan - - |
| Date Deposited: | 14 Sep 2024 13:37 |
| Last Modified: | 14 Sep 2024 13:37 |
| URI: | http://repository.stfi.ac.id/id/eprint/1206 |
Actions (login required)
 |
View Item |
