Application of T vector cloning in construction of expression vector for virus-like particles

Objective To improve the efficiency of constructing expression vector for PCR fragment with some restriction endonuclease such as Hind Ⅲ site Methods The recombinant vector, in which the PCR fragments with Hind Ⅲ site were ligated with T vector, was transformed into the BL21 DE3 E Coli Then, the recombinant plasmid vectors, which were be purified from the bacteria after its proliferation, were digested by Hind Ⅲ Finally, the target fragments with Hind Ⅲ site which purified from agarose gel after electrophoresis of the vectors having been digested by Hind Ⅲ were connected with the pNCCL1 expression vector that have been digested by the same restriction endonuclease Results The construction of expression vector for virus like particles containing SARS CoV partial RNA was succeed by use of T vector model, and the operation was simple and time saving In contrast, expression vector was not obtained by digesting the PCR products with Hind Ⅲ site directly with Hind Ⅲ Conclusion For PCR product with restriction endonuclease site, such as Hind Ⅲ site, it is a efficient and simple method way to construct a recombinant expression vector by use of T vector cloning