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You searched for subject:(pET 15b). Showing records 1 – 2 of 2 total matches.

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Linköping University

1. Lundén, Amanda. Can Sterol Carrie Protein-2 function as a solubility tag in E.coli?.

Degree: Biology, 2016, Linköping University

Expressing foreign proteins in E.coli is a major challenge because they often tend to develop into unsolvable and inactive proteins. They aggregate into so called  inclusion bodies which prevent expression of the protein. This problem might be avoided by fusing the gene of the foreign protein with a soluble protein called solubility tags, which  function is to enhance the solubility of the foreign protein. This report investigates whether Sterol Carrier Protein-2 (SCP-2) could function as a solubility tag. The experiment was carried out by fusing SCP-2 to two recombinant proteins, Green fluorescent protein (GFP) and a form of chloroamphenicol acetyl transferase (CATΔ9). The gene fusion was then inserted into a pET-15 vector and transformed into  the E.coli strain BL21(DE3) to be expressed. The results obtained from Western blot and PageBlue staining indicates that SCP-2 does not enhance the solubility of GFP or CATΔ9 since neither of them was expressed.  Furthermore, previous studies have shown that GFP can in fact be expressed  usingmaltose binding protein (MBP) as a solubility tag. Unfortunately, no success has been made regarding CATΔ9. In conclusion, regarding the results from this report, SCP-2 does not function as a solubility tag. However, further studies should be carried out on SCP-2 with more experiments before rejecting the possibility to use SCP-2 as a solubility tag.

Subjects/Keywords: CATΔ9; E.coli; GFP; pET-15b; protein expression; recombinant proteins; SCP-2; Solubility tag

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Lundén, A. (2016). Can Sterol Carrie Protein-2 function as a solubility tag in E.coli?. (Thesis). Linköping University. Retrieved from http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-129803

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Lundén, Amanda. “Can Sterol Carrie Protein-2 function as a solubility tag in E.coli?.” 2016. Thesis, Linköping University. Accessed May 09, 2021. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-129803.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Lundén, Amanda. “Can Sterol Carrie Protein-2 function as a solubility tag in E.coli?.” 2016. Web. 09 May 2021.

Vancouver:

Lundén A. Can Sterol Carrie Protein-2 function as a solubility tag in E.coli?. [Internet] [Thesis]. Linköping University; 2016. [cited 2021 May 09]. Available from: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-129803.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Lundén A. Can Sterol Carrie Protein-2 function as a solubility tag in E.coli?. [Thesis]. Linköping University; 2016. Available from: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-129803

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation


Linköping University

2. Mikkola, Isak. Does SCP-2 promote the expression of foreign proteins in Escherichia coli?.

Degree: Biology, 2016, Linköping University

Expression of foreign proteins in host organisms usually results in the development of insoluble, inactive proteins. Further, these proteins have a tendency to form aggregates termed inclusion bodies. However, the formation of inclusion bodies can be avoided by fusing the gene encoding the foreign protein to a highly soluble protein. In this report Sterol Carrier Protein-2 (SCP-2) is reviewed as a possible solubility tag. The experiment was carried out by fusing SCP-2 to one of two i nsoluble proteins, Green fluorescent protein (GFP) or a form of chloramphenicol acetyl transferase (CAT∆9). The protein fusion was then inserted into the vector pET-15b, transformed in Escherichia coli and the yield of actively expressed protein was measured. The results obtained from this study, as evaluated by PageBlue staining and  Western blot, are indicating that SCP-2 does not improve the solubility of GFP or CAT∆9. Nonetheless, the solubility of GFP has earlier been increased by fusing it to the solubility tag maltose-binding protein (MBP).  Producing more soluble forms of CAT∆9  have also been tested but without success. Therefore the conclusion drawn from this experiment is that SCP-2 does not work as a solubility tag, however more research must be performed to conclude this with certainty.

Subjects/Keywords: CAT∆9; Escherichia coli; GFP; inclusion body; pET-15b; SCP-2; solubility tag

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Mikkola, I. (2016). Does SCP-2 promote the expression of foreign proteins in Escherichia coli?. (Thesis). Linköping University. Retrieved from http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-129802

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Mikkola, Isak. “Does SCP-2 promote the expression of foreign proteins in Escherichia coli?.” 2016. Thesis, Linköping University. Accessed May 09, 2021. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-129802.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Mikkola, Isak. “Does SCP-2 promote the expression of foreign proteins in Escherichia coli?.” 2016. Web. 09 May 2021.

Vancouver:

Mikkola I. Does SCP-2 promote the expression of foreign proteins in Escherichia coli?. [Internet] [Thesis]. Linköping University; 2016. [cited 2021 May 09]. Available from: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-129802.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Mikkola I. Does SCP-2 promote the expression of foreign proteins in Escherichia coli?. [Thesis]. Linköping University; 2016. Available from: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-129802

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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