Difference between revisions of "Part:BBa K3257042"
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This part is the gene encoding capsid protein(gag) and reverse transcriptase(pol), which is the key of our mutant library construction system. The reverse transcriptase from MMLV(Moloney Murine Leukemia virus) reverse transcribe with a relative lower fidelity so that it introduces random mutations in the cDNA. In wild type Moloney Murine Leukemia virus , there is a stop codon between mRNA transcribed from the gag gene and pol gene while it can be read through as a Glutamine when translated by ribosome to form gag and gag-pol polyprotein at a ratio of 20:1. We have changed the stop codon into a Glutamine codon to assure its readability in E.coli. In the future, this part can be used to express gag-pol polyprotein inside the bacteria to initiate the reverse transcription process in vivo. | This part is the gene encoding capsid protein(gag) and reverse transcriptase(pol), which is the key of our mutant library construction system. The reverse transcriptase from MMLV(Moloney Murine Leukemia virus) reverse transcribe with a relative lower fidelity so that it introduces random mutations in the cDNA. In wild type Moloney Murine Leukemia virus , there is a stop codon between mRNA transcribed from the gag gene and pol gene while it can be read through as a Glutamine when translated by ribosome to form gag and gag-pol polyprotein at a ratio of 20:1. We have changed the stop codon into a Glutamine codon to assure its readability in E.coli. In the future, this part can be used to express gag-pol polyprotein inside the bacteria to initiate the reverse transcription process in vivo. | ||
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Revision as of 11:22, 9 September 2019
MMLV gag-pol Polyprotein
This part is the gene encoding capsid protein(gag) and reverse transcriptase(pol), which is the key of our mutant library construction system. The reverse transcriptase from MMLV(Moloney Murine Leukemia virus) reverse transcribe with a relative lower fidelity so that it introduces random mutations in the cDNA. In wild type Moloney Murine Leukemia virus , there is a stop codon between mRNA transcribed from the gag gene and pol gene while it can be read through as a Glutamine when translated by ribosome to form gag and gag-pol polyprotein at a ratio of 20:1. We have changed the stop codon into a Glutamine codon to assure its readability in E.coli. In the future, this part can be used to express gag-pol polyprotein inside the bacteria to initiate the reverse transcription process in vivo.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 174
Illegal BglII site found at 1286
Illegal BglII site found at 2592
Illegal BglII site found at 3824
Illegal BamHI site found at 2609
Illegal BamHI site found at 2915
Illegal XhoI site found at 940 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1825
Illegal AgeI site found at 3396 - 1000COMPATIBLE WITH RFC[1000]