Difference between revisions of "Part:BBa K3020002"
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The green fluorescent protein consists of about 238 amino acids, about 25KDa, which is excited by blue or ultraviolet light to produce green fluorescence. In the field of biotechnology, green fluorescent protein is often used as a reporter gene, and has a wide range of applications in molecular detection, screening, biosensors, and disease diagnosis. | The green fluorescent protein consists of about 238 amino acids, about 25KDa, which is excited by blue or ultraviolet light to produce green fluorescence. In the field of biotechnology, green fluorescent protein is often used as a reporter gene, and has a wide range of applications in molecular detection, screening, biosensors, and disease diagnosis. | ||
The reporter gene used in this experiment is a mutant of GFP, which enhances green fluorescent protein and its fluorescence intensity is more than 6 times that of GFP. It is more suitable as a reporter factor for gene expression, regulation and protein localization and transport than GFP. The eGFP gene sequence used in this experiment was obtained from NCBI, sequence ID: 20473140. Due to the preference of codons, the level of exogenous gene expression was affected. The gene sequence was submitted to Jin Weizhi (Suzhou) Bio Co., Ltd. for codon optimization of E. coli host. The 5'-3' end of the optimized eGFP sequence was synthesized between the XbaI and HindIII restriction sites in the multiple cloning site of pUC19 plasmid. The plasmid map is shown below. | The reporter gene used in this experiment is a mutant of GFP, which enhances green fluorescent protein and its fluorescence intensity is more than 6 times that of GFP. It is more suitable as a reporter factor for gene expression, regulation and protein localization and transport than GFP. The eGFP gene sequence used in this experiment was obtained from NCBI, sequence ID: 20473140. Due to the preference of codons, the level of exogenous gene expression was affected. The gene sequence was submitted to Jin Weizhi (Suzhou) Bio Co., Ltd. for codon optimization of E. coli host. The 5'-3' end of the optimized eGFP sequence was synthesized between the XbaI and HindIII restriction sites in the multiple cloning site of pUC19 plasmid. The plasmid map is shown below. | ||
| − | + | [[File:BIT201903-puc19-egfp.png|thumb|400px|center]] | |
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===Usage and Biology=== | ===Usage and Biology=== | ||
Latest revision as of 08:29, 8 September 2019
enhanced Green fluorescent protein reporter gene
The green fluorescent protein consists of about 238 amino acids, about 25KDa, which is excited by blue or ultraviolet light to produce green fluorescence. In the field of biotechnology, green fluorescent protein is often used as a reporter gene, and has a wide range of applications in molecular detection, screening, biosensors, and disease diagnosis. The reporter gene used in this experiment is a mutant of GFP, which enhances green fluorescent protein and its fluorescence intensity is more than 6 times that of GFP. It is more suitable as a reporter factor for gene expression, regulation and protein localization and transport than GFP. The eGFP gene sequence used in this experiment was obtained from NCBI, sequence ID: 20473140. Due to the preference of codons, the level of exogenous gene expression was affected. The gene sequence was submitted to Jin Weizhi (Suzhou) Bio Co., Ltd. for codon optimization of E. coli host. The 5'-3' end of the optimized eGFP sequence was synthesized between the XbaI and HindIII restriction sites in the multiple cloning site of pUC19 plasmid. The plasmid map is shown below.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 682
- 1000COMPATIBLE WITH RFC[1000]