The invention provides a
gene synthesis method. The method comprises the steps that 1, a to-be-synthesized
gene sequence is divided into a first continuous fragment with the length being 55-100 bp, the first continuous fragment is designed to be a
forward primer, a
gene sequence staggered with the first continuous fragment at the portion of the length of 15-20 bp serves as a second continuous fragment, and continuous reverse primers are designed on a reverse complementary sequence of the second continuous fragment; 2, the
forward primer is mixed with the reverse primers; 3, the primer mixture is boiled, annealing is conducted, T4
DNA ligase is added, and the primers are connected into a double-stranded
DNA gene with the two ends provided with the long sticky
tail ends; 4, the synthesized double-stranded
DNA gene is connected to a pUC57-GFP-Amp carrier, the sequence of the pUC57-GFP-Amp carrier is shown as a sequence table 1,
escherichia coli cells are converted, a non-fluorescing
bacterial colony is reversely screened, and sequencing
verification is conducted. Accordingly, the method does not reply on PCR amplification, and the defects that the experimental procedure in PCR amplification is tedious, and the PCR technical error rate is high are overcome.