Comparison of Two Construction Methods of Targeting Gene of Haemophilus parasuis

摘要

Haemophilus parasuis is one kind of constant bacteria in porcine upper respiratory tract,and it can cause multiple serositis,arthritis and other diseases under certain conditions.Due to lack of efficient genetic operating system,its pathogenic mechanism is not very clear.Ligation with DNA ligase and fusion PCR were used to construct targeting hhdA gene of Haemophilus parasuis,respectively.The fidelity,application scope,operation and conditions of the constructed fusion fragments were compared.The results showed that construction with DNA ligase was more mature technology as manifested by more stable conditions and more extensive application.The fusion PCR method had high fidelity and simple operation,and the transformation rate was 9.5 times as high as that of ligation with DNA ligase.For this reason,this method was more suitable for construction of multi-fragment targeting genes.The study lays a foundation for establishing an efficient operating system of targeting gene of Haemophilus parasuis in the future.