Đề bài: Xây dựng vecto pHT43 sản xuất enzyme nattokinase có đuôi
His ở đầu C ở Bacillus subtilis bằng Gibson và Golden Gate
Vùng inhibitor là vùng ức chế cần loại bỏ
>D25319.1:181-1326 Bacillus natto subtilisin gene, complete cds
aTGAGAAGCAAAAAATTGTGGATCAGCTTGTTGTTTGCGTTAACGTTAATCTTTACGATGGCGTTCAGCA
ACATGTCTGCGCAGGCTGCCGGAAAAAGCAGTACAGAAAAGAAATACATTGTCGGATTTAAGCAGACAAT
GAGTGCCATGAGTTCCGCCAAGAAAAAGGATGTTATTTCTGAAAAAGGCGGAAAGGTTCAAAAGCAATTT
AAGTATGTTAACGCGGCCGCAGCAACATTGGATGAAAAAGCTGTAAAAGAATTGAAAAAAGATCCGAGCG
TTGCATATGTGGAAGAAGATCATATTGCACATGAATAT
GCGCAATCTGTTCCTTATGGCATTTCTCAAATTAAAGCGCCGGCTCTTCACTCTCAAGGCTACACAGGCTCTAACGT
AAAAGTAGCTGTTATCGACAGCGGAATTGACTCTTCTCATCCTGACTTAAACGTCAGAGGCGGAGCAAGCTTCGTTC
CTTCTGAAACAAACCCATACCAGGACGGCAGTTCTCACGGTACGCATGTCGCCGGTACGATTGCCGCTCTTAATAAC
TCAATCGGTGTTCTGGGCGTAGCGCCAAGCGCATCATTATATGCAGTAAAAGTGCTTGATTCAACAGGAAGCGGCCA
ATATAGCTGGATTATTAACGGCATTGAGTGGGCCATTTCCAACAATATGGATGTTATCAACATGAGCCTTGGCG
GACCTACTGGTTCTACAGCGCTGAAAACAGTAGTTGATAAAGCGGTTTCCAGCGGTATCGTCGTTGCTGC
CGCAGCCGGAAACGAAGGTTCATCCGGAAGCACAAGCACAGTCGGCTACCCTGCAAAATATCCTTCTACT
ATTGCAGTAGGTGCGGTAAACAGCAGCAACCAAAGAGCTTCATTCTCCAGCGTAGGTTCTGAGCTTGATG
TAATGGCTCCTGGCGTGTCCATCCAAAGCACACTTCCTGGAGGCACTTACGGCGCTTATAACGGAACGTC
CATGGCGACTCCTCACGTTGCCGGAGCAGCAGCGCTAATTCTTTCTAAGCACCCGACTTGGACAAACGCG
CAAGTCCGTGATCGTTTAGAAAGCACTGCAACATATCTTGGAAACTCTTTCTACTATGGAAAAGGGTTAA
TCAACGTACAAGCAGCTGCACAA
TAA
Đuôi His đầu C xóa mã kết thúc trong gen
Mã khởi đầu có thể là GTG,TTG => muốn biểu hiện mạnh đưa về ATG
LEFT PRIMER 1 21 60.27 52.38 0.30 0.00 0.00
GCGCAATCTGTTCCTTATGGC
RIGHT PRIMER 825 20 60.81 50.00 36.61 0.00 42.66
TTGTGCAGCTGCTTGTACGT
>pHT43 ( chèn gen vào vecto phải đảo đoạn dưới lên trên )
GTAGGATCCTCTAGAGTCGACGTCCCCGGGGCAGCCCGCCTAATGAGCGGGCTTTTTTCACGTC
ACGCGTCCATGGAGATCTTTGTCTGCAACTGAAAAGTTTATACCTTACCTGGAACAAATGGTTG
AAACATACGAGGCTAATATCGGCTTATTAGGAATAGTCCCTGTACTAATAAAATCAGGTGGATC
AGTTGATCAGTATATTTTGGACGAAGCTCGGAAAGAATTTGGAGATGACTTGCTTAATTCCACA
ATTAAATTAAGGGAAAGAATAAAGCGATTTGATGTTCAAGGAATCACGGAAGAAGATACTCATG
ATAAAGAAGCTCTAAAACTATTCAATAACCTTACAATGGAATTGATCGAAAGGGTGGAAGGTTA
ATGGTACGAAAATTAGGGGATCTACCTAGAAAGCCACAAGGCGATAGGTCAAGCTTAAAGAACC
CTTACATGGATCTTACAGATTCTGAAAGTAAAGAAACAACAGAGGTTAAACAAACAGAACCAAA
AAGAAAAAAAGCATTGTTGAAAACAATGAAAGTTGATGTTTCAATCCATAATAAGATTAAATCG
CTGCACGAAATTCTGGCAGCATCCGAAGGGAATTCATATTACTTAGAGGATACTATTGAGAGAG
CTATTGATAAGATGGTTGAGACATTACCTGAGAGCCAAAAAACTTTTTATGAATATGAATTAAA
AAAAAGAACCAACAAAGGCTGAGACAGACTCCAAACGAGTCTGTTTTTTTAAAAAAAATATTAG
GAGCATTGAATATATATTAGAGAATTAAGAAAGACATGGGAATAAAAATATTTTAAATCCAGTA
AAAATATGATAAGATTATTTCAGAATATGAAGAACTCTGTTTGTTTTTGATGAAAAAACAAACA
