褐环黏盖牛肝菌中异柠檬酸裂解酶基因的简并引物设计

Degenerate Primers Design of Suillus Luteus Isocitrate Lyase Gene

异柠檬酸裂解酶是乙醛酸支路中的关键酶,研究褐环黏盖牛肝菌中异柠檬酸裂解酶的基因序列,能为今后研究其结构和性质提供科学依据。关于未知序列引物的设计,使用NCBI,Uni Prot,PROSITE,Blast等生物信息资源和DNAMAN,Oligo 6.0等生物软件,结合异柠檬酸裂解酶的催化模体和密码子的偏好性,设计褐环黏盖牛肝菌中异柠檬酸裂解酶的简并引物。简并引物的成功设计,对后续褐环黏盖牛肝菌中异柠檬酸裂解酶基因的克隆奠定基础。结果表明,简并引物的设计能够提高引物的特异性,有利于试验成功。

Isocitrate lyase is the key enzyme of glyoxalic acid branch. Researching the gene sequences of isocitrate lyase in the Suillus luteus can provide the scientific basis for future studies of its structure and properties. This article explores about the design of the unknown sequence primers. Used NCBI, UniProt, PROSITE, Blast and other biological information resources and DNAMAN, Oligo 6.0 biological software, and combined with isocitrate lyase catalytic motif and bias of codon, to design Suillus luteus isocitrate lyase degenerate primers. The successful design of the degenerate primers lay the foundation for the subsequent cloning of Suillus luteus isocitrate lyase gene. The results show that the design of degenerate primers can improve the specificity of the primers, which is helpful to the successful of the experiments.