Difference between revisions of "Part:BBa K1694010"
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<h1>'''Introduction'''</h1> | <h1>'''Introduction'''</h1> | ||
| − | Lpp-OmpA protein is | + | Lpp-OmpA protein is the expression system of outer membrane protein, which consists of the 20 amino acids of signal sequence, the 9 N-terminal amino acids of the lipoprotein (Lpp) and the residual 46-159 amino acids of the OmpA.<br> |
The lipoprotein (Lpp) is the most abundant protein on the outer membrane, which possesses the function of targeting to the outer membrane. <br> | The lipoprotein (Lpp) is the most abundant protein on the outer membrane, which possesses the function of targeting to the outer membrane. <br> | ||
| − | The OmpA domain constitutes eight-stranded β-barrel to construct an anchor on the outer membrane, providing the stable expression of the | + | The OmpA domain constitutes eight-stranded β-barrel to construct an anchor on the outer membrane, providing the stable expression of the protein on the outer membrane.<br> |
| − | By taking advantage of | + | By taking advantage of efficiently targeting OmpA to the outer membrane, it allows C-terminal fusion of the scFv sequence to be displayed out of the outer mambrane.<br> |
<h2>'''Experiment'''</h2> | <h2>'''Experiment'''</h2> | ||
| − | After PCR the DNA sequences from Lpp-OmpA-N , we | + | After PCR the DNA sequences from Lpp-OmpA-N , we got the Lpp-OmpA, and then recombined each Lpp-OmpA gene with PSB1C3 backbone and conducted a PCR experiment to check the size of each of the Lpp-OmpA. The DNA sequence length of the Lpp-OmpA is around 400~500 bp. In this PCR experiment, the Lpp-OmpA products size should be near at 650~750 bp. The Fig. showed the correct size of the Lpp-OmpA, and proved that we successfully ligated the Lpp-OmpA sequence onto an ideal backbone. |
| − | [[File:OmpA.png|200px|thumb|left|The DNA sequence length of the Lpp-OmpA is around 400~500 bp. In this PCR experiment, the Lpp-OmpA products size should be near at 650~750 bp.]] | + | [[File:Lpp-OmpA.png|200px|thumb|left|The DNA sequence length of the Lpp-OmpA is around 400~500 bp. In this PCR experiment, the Lpp-OmpA products size should be near at 650~750 bp.]] |
Latest revision as of 21:42, 22 September 2015
Lpp-OmpA
Introduction
Lpp-OmpA protein is the expression system of outer membrane protein, which consists of the 20 amino acids of signal sequence, the 9 N-terminal amino acids of the lipoprotein (Lpp) and the residual 46-159 amino acids of the OmpA.
The lipoprotein (Lpp) is the most abundant protein on the outer membrane, which possesses the function of targeting to the outer membrane.
The OmpA domain constitutes eight-stranded β-barrel to construct an anchor on the outer membrane, providing the stable expression of the protein on the outer membrane.
By taking advantage of efficiently targeting OmpA to the outer membrane, it allows C-terminal fusion of the scFv sequence to be displayed out of the outer mambrane.
Experiment
After PCR the DNA sequences from Lpp-OmpA-N , we got the Lpp-OmpA, and then recombined each Lpp-OmpA gene with PSB1C3 backbone and conducted a PCR experiment to check the size of each of the Lpp-OmpA. The DNA sequence length of the Lpp-OmpA is around 400~500 bp. In this PCR experiment, the Lpp-OmpA products size should be near at 650~750 bp. The Fig. showed the correct size of the Lpp-OmpA, and proved that we successfully ligated the Lpp-OmpA sequence onto an ideal backbone.
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 388
- 1000COMPATIBLE WITH RFC[1000]