Fosmid T14B1.2 containing aex-2
sequence was transformed into recombineering strain SW102. A galK gene was inserted into the C -terminus of the aex-2
coding region with following oligonucleotides: forward GACGAGCATCTGAAAGGCCGCCGGAGCACACCCCCTTACGGTGTGATATGCCTGTTGACAATTAATCATCGGCA, reverse CATTTTTTCCACAAGTTTTACTTACATACATTGCGAATTACTACGATCTATCAGCACTGTCCTGCTCCTT. The galK gene was subsequently replaced by the mCherry gene by homologous recombination with the following oligonucleotides: forward GACGAGCATCTGAAAGGCCGCCGGAGCACACCCCCTTACGGTGTGATATGATGGTGAGCAAGGGCGAGGAG, reverse CATTTTTTCCACAAGTTTTACTTACATACATTGCGAATTACTACGATCTACTTGTACAGCTCGTCCATGCC. In the final step, the whole fragment, which contains aex-2
coding sequence, mCherry coding sequence, and 3'-UTR, was gap repaired into an Amp containing vector backbone using oligonucleotides for ward CATTGATCTGCCGCATGATGAAGTACCAAGTCTGAATGATGAAGAATTTCATTCGTTATGCATTATGGGTAC and reverse AATCAAACGACATTAACGATTTCTCAAAAAAAAAAAACTTTAGGAAAACATACCAATCTAAGTCTGTGCTCC and was used to make transgenes jsEx937 and jsEx938. --precise ends.