Table 2.
Open in new tab

Primers used for the construction of hybrid genes and of mutated cDNAs

cDNA cloneTemplateManipulationForward primerReverse primer
Hybrid genesORF region
C15′-fragment: F3′H1–196ATGACTATTCTACCCCTACTACTCCTTCGCATCACGCTCACCG
3′-fragment: CH3H197–508P-GCGGATGAGTTCAAGGATATGGCCTTAATGACTTCCATACACGTG
C25′-fragment: CH3H1–210ATGACTATTCTACCCCTACTACTCCTTCGGATCACCACCTGCA
3′-fragment: F3'H211–512P-GCGGATGAGTTCAAG GAT ATG GAGGATGTCTCAACTTACAACAATAAACC
C35′-fragment: CH3H1–119ATGACTATTCTACCCCTACTACTCGTACGGTGCAAACACCATATC
3′-fragment: F3′H120–508P- GGTCCTCGGTGGCGGATAGGATGTCTCAACTTACAACAATAAACC
C45′-fragment: CH3H1–193ATGACTATTCTACCCCTACTACTCP- TTCGCATCTCCGTCGGC
3′-fragment: F3′H194–508GGCGGATGAGTTCAAGGATATGAGGATGTCTCAACTTACAACAATAAACC
C55′-fragment: CH3H1–119ATGACTATTCTACCCCTACTACTCGTACGGTGCAAACACCATATC
3′-fragment: M2120–512P- GGTCCTCGGTGGCGGATAGGATGTCTCAACTTACAACAATAAACC
Mutated genesMutation
M15′-fragment: CH3HDeletion of S(194)AAG(197)ATGACTATTCTACCCCTACTACTCTCCGTCGGCGAATACTCTC
3′-fragment: CH3HP-GATCCGAAGGCGGATGAGTTCCTTAATGACTTCCATACACGTG
M25′-fragment F3′HInsertion of S(194)AAG(197), exchange of R(193) into G and A(199) into PATGACTATTCTACCCCTACTACTCCACCTGCACTTCCGTCACCGAATACTCTCCTGCC (SAGG from CH3H/F3′H)
3′-fragment: F3′HP-GTGATGCGAAGGCGGATGAGAGGATGTCTCAACTTACAACAATAAACC
M3CH3HCCG -> GCG P(199) -> AATCCGCCTTCGCATCACCA
cDNA cloneTemplateManipulationForward primerReverse primer
Hybrid genesORF region
C15′-fragment: F3′H1–196ATGACTATTCTACCCCTACTACTCCTTCGCATCACGCTCACCG
3′-fragment: CH3H197–508P-GCGGATGAGTTCAAGGATATGGCCTTAATGACTTCCATACACGTG
C25′-fragment: CH3H1–210ATGACTATTCTACCCCTACTACTCCTTCGGATCACCACCTGCA
3′-fragment: F3'H211–512P-GCGGATGAGTTCAAG GAT ATG GAGGATGTCTCAACTTACAACAATAAACC
C35′-fragment: CH3H1–119ATGACTATTCTACCCCTACTACTCGTACGGTGCAAACACCATATC
3′-fragment: F3′H120–508P- GGTCCTCGGTGGCGGATAGGATGTCTCAACTTACAACAATAAACC
C45′-fragment: CH3H1–193ATGACTATTCTACCCCTACTACTCP- TTCGCATCTCCGTCGGC
3′-fragment: F3′H194–508GGCGGATGAGTTCAAGGATATGAGGATGTCTCAACTTACAACAATAAACC
C55′-fragment: CH3H1–119ATGACTATTCTACCCCTACTACTCGTACGGTGCAAACACCATATC
3′-fragment: M2120–512P- GGTCCTCGGTGGCGGATAGGATGTCTCAACTTACAACAATAAACC
Mutated genesMutation
M15′-fragment: CH3HDeletion of S(194)AAG(197)ATGACTATTCTACCCCTACTACTCTCCGTCGGCGAATACTCTC
3′-fragment: CH3HP-GATCCGAAGGCGGATGAGTTCCTTAATGACTTCCATACACGTG
M25′-fragment F3′HInsertion of S(194)AAG(197), exchange of R(193) into G and A(199) into PATGACTATTCTACCCCTACTACTCCACCTGCACTTCCGTCACCGAATACTCTCCTGCC (SAGG from CH3H/F3′H)
3′-fragment: F3′HP-GTGATGCGAAGGCGGATGAGAGGATGTCTCAACTTACAACAATAAACC
M3CH3HCCG -> GCG P(199) -> AATCCGCCTTCGCATCACCA
Table 2.
Open in new tab

Primers used for the construction of hybrid genes and of mutated cDNAs

cDNA cloneTemplateManipulationForward primerReverse primer
Hybrid genesORF region
C15′-fragment: F3′H1–196ATGACTATTCTACCCCTACTACTCCTTCGCATCACGCTCACCG
3′-fragment: CH3H197–508P-GCGGATGAGTTCAAGGATATGGCCTTAATGACTTCCATACACGTG
C25′-fragment: CH3H1–210ATGACTATTCTACCCCTACTACTCCTTCGGATCACCACCTGCA
3′-fragment: F3'H211–512P-GCGGATGAGTTCAAG GAT ATG GAGGATGTCTCAACTTACAACAATAAACC
C35′-fragment: CH3H1–119ATGACTATTCTACCCCTACTACTCGTACGGTGCAAACACCATATC
3′-fragment: F3′H120–508P- GGTCCTCGGTGGCGGATAGGATGTCTCAACTTACAACAATAAACC
C45′-fragment: CH3H1–193ATGACTATTCTACCCCTACTACTCP- TTCGCATCTCCGTCGGC
3′-fragment: F3′H194–508GGCGGATGAGTTCAAGGATATGAGGATGTCTCAACTTACAACAATAAACC
C55′-fragment: CH3H1–119ATGACTATTCTACCCCTACTACTCGTACGGTGCAAACACCATATC
3′-fragment: M2120–512P- GGTCCTCGGTGGCGGATAGGATGTCTCAACTTACAACAATAAACC
Mutated genesMutation
M15′-fragment: CH3HDeletion of S(194)AAG(197)ATGACTATTCTACCCCTACTACTCTCCGTCGGCGAATACTCTC
3′-fragment: CH3HP-GATCCGAAGGCGGATGAGTTCCTTAATGACTTCCATACACGTG
M25′-fragment F3′HInsertion of S(194)AAG(197), exchange of R(193) into G and A(199) into PATGACTATTCTACCCCTACTACTCCACCTGCACTTCCGTCACCGAATACTCTCCTGCC (SAGG from CH3H/F3′H)
3′-fragment: F3′HP-GTGATGCGAAGGCGGATGAGAGGATGTCTCAACTTACAACAATAAACC
M3CH3HCCG -> GCG P(199) -> AATCCGCCTTCGCATCACCA
cDNA cloneTemplateManipulationForward primerReverse primer
Hybrid genesORF region
C15′-fragment: F3′H1–196ATGACTATTCTACCCCTACTACTCCTTCGCATCACGCTCACCG
3′-fragment: CH3H197–508P-GCGGATGAGTTCAAGGATATGGCCTTAATGACTTCCATACACGTG
C25′-fragment: CH3H1–210ATGACTATTCTACCCCTACTACTCCTTCGGATCACCACCTGCA
3′-fragment: F3'H211–512P-GCGGATGAGTTCAAG GAT ATG GAGGATGTCTCAACTTACAACAATAAACC
C35′-fragment: CH3H1–119ATGACTATTCTACCCCTACTACTCGTACGGTGCAAACACCATATC
3′-fragment: F3′H120–508P- GGTCCTCGGTGGCGGATAGGATGTCTCAACTTACAACAATAAACC
C45′-fragment: CH3H1–193ATGACTATTCTACCCCTACTACTCP- TTCGCATCTCCGTCGGC
3′-fragment: F3′H194–508GGCGGATGAGTTCAAGGATATGAGGATGTCTCAACTTACAACAATAAACC
C55′-fragment: CH3H1–119ATGACTATTCTACCCCTACTACTCGTACGGTGCAAACACCATATC
3′-fragment: M2120–512P- GGTCCTCGGTGGCGGATAGGATGTCTCAACTTACAACAATAAACC
Mutated genesMutation
M15′-fragment: CH3HDeletion of S(194)AAG(197)ATGACTATTCTACCCCTACTACTCTCCGTCGGCGAATACTCTC
3′-fragment: CH3HP-GATCCGAAGGCGGATGAGTTCCTTAATGACTTCCATACACGTG
M25′-fragment F3′HInsertion of S(194)AAG(197), exchange of R(193) into G and A(199) into PATGACTATTCTACCCCTACTACTCCACCTGCACTTCCGTCACCGAATACTCTCCTGCC (SAGG from CH3H/F3′H)
3′-fragment: F3′HP-GTGATGCGAAGGCGGATGAGAGGATGTCTCAACTTACAACAATAAACC
M3CH3HCCG -> GCG P(199) -> AATCCGCCTTCGCATCACCA
Close
This Feature Is Available To Subscribers Only

Sign In or Create an Account

Close

This PDF is available to Subscribers Only

View Article Abstract & Purchase Options

For full access to this pdf, sign in to an existing account, or purchase an annual subscription.

Close