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The Analytical and Purification Methodology to Confirm the Presence of a GFP Gene in the Expression Plasmid pet28a Using 6x Histidine-Tag
| المصدر: | عالم التربية |
|---|---|
| الناشر: | المؤسسة العربية للاستشارات العلمية وتنمية الموارد البشرية |
| المؤلف الرئيسي: | AlSarraj, Faisal (Author) |
| المجلد/العدد: | س17, ع54 |
| محكمة: | نعم |
| الدولة: |
مصر |
| التاريخ الميلادي: |
2016
|
| الشهر: | ابريل |
| الصفحات: | 1 - 17 |
| DOI: |
10.12816/0031906 |
| ISSN: |
1110-4406 |
| رقم MD: | 826312 |
| نوع المحتوى: | بحوث ومقالات |
| اللغة: | الإنجليزية |
| قواعد المعلومات: | EduSearch |
| مواضيع: | |
| رابط المحتوى: |
عدد مرات التحميل
12
| المستخلص: |
The field of molecular biology is quickly advancing. New discoveries are being made that greatly enhance the field. One such discovery is the green fluorescent protein. This discovery has revolutionized the research being done in the field of molecular biology. The green fluorescent protein is used to track, visualize and quantify the gene derivatives in living tissues. The aim of this study was to prove the existence of a 6xhistidine-tagged GFP gene in the plasmid pET28a. Earlier, the gene was inserted in the plasmid and had to be confirmed. The procedures used in obtaining the results were mainly purification techniques that included chromatography, ion-exchange and immobilised metal affinity chromatography or IMAC. The DNA-fragments were analysed using the Aragose gel electrophoresis technique, after which the SDS-PAGE analysis was carried out. The size of the resultant GFP that was obtained from the plasmid pET28a was 1200bp. After the application of numerous purification steps, the sample that was acquired was of a high quality and the results showed that it had a molecular weight of approximately 28- 30 kDa. After these purification steps, it was analysed using the SDS-PAGE technique. |
|---|---|
| ISSN: |
1110-4406 |
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