Improved overlap extension PCR process and mutation gene obtained thereby
A technology of overlapping extension and gene mutation, applied in fermentation and other directions, can solve the problems of cumbersome test steps and heavy workload, and achieve the effects of increasing accumulation, good fidelity and high-efficiency expression
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Embodiment 1
[0032] A gene sam1 encoding SAM synthetase (2.5.1.6) derived from Saccharomyces cerevisiae INVScI strain has the base sequence in SEQ ID No: 1 in the sequence table.
[0033] SEQ ID No: 1:
[0034] ATGGCCGGTACATTTTTATTCACTTCTGAATCCGTTGGTGAAGGTCACCCGAGA
[0035] TAAGATCTGTGACCAAGTTTCCGACGCCATCTTGGACGCTTGTTTAGCCGAGG
[0036] ACCCTCACTCCAAAGTTGCGTGTGAAACCGCGGCAAAGACTGGTATGATTATG
[0037] GTCTTTGGTGAAATTACTACCAAGGCACAGTTGGATTACCAAAAAATCGTCAG
[0038] AGACACCATCAAGAAGATTGGTTACGATGATTCCGCCAAGGGTTTCGACTATA
[0039] AGACCTGTAACGTCCTTGTCGCCATTGAGCAACAATCTCCAGATATCGCCCAA
[0040] GGTGTCCACGAGGAGAAGGATTTGGAAGACATCGGTGCCGGTGACCAAGGTAT
[0041] CATGTTTGGTTACGCCACAGATGAAACTCCAGAGGGTTTGCCTTTGACTATTC
[0042] TTTTGGCTCATAAACTAAACATGGCCATGGCTGACGCGAGAAGAGATGGCTCT
[0043] TTAGCGTGGTTGAGACCAGACACCAAGACTCAAGTCACCGTCGAATACAAGGA
[0044] TGACCACGGTAGATGGGTTCCACAAAAGAATCGACACCGTCGTCGTCTCCGCTC
[0045] AACATGCTGACGAAATCACGACCGAGGACTTAAGAGCGCAACTAAAGTCCGAG
[0046] ATCATTGAAAAAGTCATCCCAAGAG...
Embodiment 2
[0072] The sam1 is shuffled by using a new comprehensive gene shuffling method to obtain the gene sam1 encoding a recombinase with improved enzyme activity, which has the base sequence of SEQ ID No: 3 in the sequence table.
[0073] ATGGCCGGTACATTTTTATTCACTTCTGAATCCGTTGGTGAAGGTCACCCGAGA
[0074] TAAGCTCTGTGACCAAGTTTCCGACGCCATCTTGGACGCTTGTTTAGCCGAGG
[0075] ACCCTCACTCCAAAGTTGCTTGTGAAACCGCTGCAAAGACTGGTATGATTATG
[0076] GTCTTTGGTGAAATTACTACCAAGGCACAGTTGGATTACCAAAAAATCGTCAG
[0077] AGACACCATCAAGAAGATTGGTTACGATGATTCCGCCAAGGGATTCGACTATA
[0078] AGACCTGTAACGTCCTTGTCGCCATTGAGCAACAATCTCCAGACATCGCCCAA
[0079] GGTGTCCACGAGGAGAAGGATTTGGAAGACATCGGTGCCAGTGACCAAGGTAT
[0080] CATGTTTGGTTACGCCACAGATGAAACTCCAGAGGGTTTGCCTTTAACTATTC
[0081] TTTTGGCTCATAAACTAAACATGGCCATGGCTGACGCTAGAAGAGATGGCTCT
[0082] TTAGCTTGGTTGAGACCAGACACCAAGACTCAAGTCACCGTCGAATACAAGGA
[0083] TGACCACGGTAGATGGGTTCCGCAAAGAATCGACACCGTCGTCGTCTCCGCTC
[0084] AACATGCTGACGAAATCACTACCGAGGACTTAAGAGCTCAACTAAAAGTCCGAG
...
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