| 摘要 |
<p>Disclosed is a method of constructing a transgene or a method for simultaneously synthesizing an array of transgenes the method comprising the steps of: a. providing a cloning vector plasmid with a backbone comprising first and second docking points capable of accepting a sequential arrangement of inserts, each docking point being fixed within the backbone and comprising at least one non-variable rare endonuclease site for an endonuclease enzyme; b. providing at least a first insert and a second insert to be included in the transgene, each insert comprising a 5' end, a nucleotide sequence of interest and a 3' end, wherein the 5' end of the first insert is compatible to the 3' end of the cleaved first docking point, and the 3' end of the second insert is compatible to the 5' end of the cleaved second docking point; and c. transferring both the first insert and the second insert into the backbone in a single reaction. Further discloses is an alternative method of constructing a transgene or a method for simultaneously synthesizing an array of transgenes the method comprising the steps of: a. providing a cloning vector plasmid comprising first and second docking points; b. introducing first nucleotide sequences to be included in the transgene into a first shuttle vector; c. introducing second nucleotide sequences to be included in the transgene into a second shuttle vector; and d. transferring simultaneously the first nucleotide sequences and the second nucleotide sequences from the shuttle vectors to the cloning vector plasmid, between the first and second docking points, wherein the 5' end of the first insert is compatible to the 3' end of the first docking point, and the 3' end of the second insert is compatible to the 5' end of the second docking point.</p> |
