OPTIMASI EKSPRESI PROTEIN NANOBODI KORTISOL PADA BAKTERI Escherichia coli BL21(DE3) DENGAN VARIASI MEDIA PERTUMBUHAN

ELITA AMELIA APRIANTI, - (2023) OPTIMASI EKSPRESI PROTEIN NANOBODI KORTISOL PADA BAKTERI Escherichia coli BL21(DE3) DENGAN VARIASI MEDIA PERTUMBUHAN. Skripsi thesis, Sekolah Tinggi Farmasi Indonesia.

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Abstract

Nanobodi, sebagai fragmen antibodi, memiliki ukuran nanometer yang memungkinkannya menembus jaringan dan sel dengan mudah. Dalam penelitian ini, ekspresi protein nanobodi kortisol dieksplorasi pada bakteri Escherichia coli BL21(DE3) dengan menggunakan media pertumbuhan. Penelitian ini bertujuan untuk memilih media pertumbuhan yang optimal untuk ekspresi nanobodi kortisol pada bakteri E.coli BL21 (DE3). Plasmid pET28a dengan nanobodi natif dan mutan dimasukkan ke dalam bakteri menggunakan kejut panas. Dua jenis media, Luria Bertani (LB) dan Nutrient Broth (NB) digunakan dengan waktu inkubasi 18 jam. Elektroforesis gel agarosa untuk memvalidasi DNA Plasmid sedangkan SDS PAGE digunakan untuk menganalisis protein berdasarkan berat molekul. Hasil menunjukkan bahwa variasi media pertumbuhan LB dan NB tidak memiliki perbedaan terhadap berat sel bakteri transforman setelah produksi, baik pada natif (0,402 mg) maupun mutan (0,424 mg). Hasil juga menunjukkan bahwa tidak terdapat perbedaan produksi protein pada nanobodi mutan dan nanobodi natif pada masing-masing media pertumbuhan, LB dan NB . Hasil SDS PAGE menunjukkan nanobodi natif menghasilkan pita protein lebih tebal daripada nanobodi mutan, yaitu Area Under the Curve (AUC) nanobodi natif (9270.4272 piksel) lebih tinggi dibandingkan AUC nanobodi mutan (74352.280 piksel) pada variasi media pertumbuhan. Disimpulkan bahwa variasi media yang digunakan tidak memiliki perbedaan yang signifikan terhadap berat sel bakteri yang dihasilkan pada kondisi eksperimen yang diuji. --- Nanobodies, as antibody fragments, have a nanometer size that allows them to penetrate tissues and cells easily. In this study, the expression of the cortisol nanobody protein was explored in Escherichia coli BL21(DE3) using growth media. This study aims to select the optimal growth medium for the expression of cortisol nanobodies in E.coli BL21 (DE3) bacteria. Plasmid pET28a with native and mutant nanobodies was introduced into the bacteria using heat shock. Two types of media, Luria-Bertani (LB) and Nutrient Broth (NB) were used with an incubation time of 18 hours. Agarose gel electrophoresis to validate Plasmid DNA while SDS-PAGE was used to analyze proteins based on molecular weight. The results showed that the variation of LB and NB growth media had no difference in the cell weight of transformant bacteria after production, both in native (0.402 mg) and mutant (0.424 mg). The results also showed that there was no difference in protein production in the mutant and native nanobodies on each growth medium, LB and NB. SDS PAGE results showed that native nanobodies produced thicker protein bands than mutant nanobodies, namely the Area Under the Curve (AUC) of native nanobodies (9270,4272 pixels) was higher than AUC of mutant nanobodies (74,352,280 pixels) in various growth media. It was concluded that the variety of media used did not have a significant difference in the weight of the bacterial cells produced under the experimental conditions tested.

Item Type:	Thesis (Skripsi)
Uncontrolled Keywords:	Nanobodi, Media Pertumbuhan, kortisol, Luria Bertani (LB) dan Nutrient Broth (NB) E.coli BL21(DE3). Nanobodies, Growth Media, cortisol, Luria Bertani (LB) and Nutrient Broth (NB) E.coli BL21(DE3)
Subjects:	Q Science > Q Science (General) Q Science > QD Chemistry
Divisions:	Program Studi S1 Farmasi
Depositing User:	pustakawan - -
Date Deposited:	06 Jun 2024 06:02
Last Modified:	06 Jun 2024 06:06
URI:	http://repository.stfi.ac.id/id/eprint/175

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