Kusano M, Shibuya M, Sankawa U, Ebizuka Y
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Biol Pharm Bull. 1995 Jan;18(1):195-7. doi: 10.1248/bpb.18.195.
A cDNA encoding rat 2,3-oxidosqualene: lanosterol cyclase, the enzyme responsible for the complex cyclization/rearrangement step in sterol biosynthesis, was cloned by extensive application of PCRs. Oligonucleotide primers were designed in sense and anti-sense directions corresponding to internal peptide sequences of purified enzyme. Successive PCRs with all possible combinations of these primers yielded a 178-bp fragment, and based on its sequence full nucleotide sequence of cDNA was obtained by a "rapid amplification of cDNA ends" (RACE) method.
编码大鼠2,3-氧化角鲨烯：羊毛甾醇环化酶（该酶负责甾醇生物合成中复杂的环化/重排步骤）的cDNA通过广泛应用聚合酶链反应（PCR）进行克隆。根据纯化酶的内部肽段序列，设计了正向和反向寡核苷酸引物。这些引物的所有可能组合进行连续PCR，得到一个178bp的片段，并基于其序列通过“cDNA末端快速扩增”（RACE）方法获得了cDNA的完整核苷酸序列。
