摘要
根据锤头状核酶(Ribozyme)的作用模式,设计、合成并克隆了特异性切割12脂加氧酶(12LO)mRNA的核酶基因。以合成的25个核苷酸长的12脂加氧酶RNA片段为底物与转录的核酶RNA一起保温检测其体外切割活性。实验结果表明,在37℃保温时,核酶在体外对12脂加氧酶具有较高的特异切割活性,其Km值为1300nmol/L,其kcat值为0083/min,在50℃保温时,核酶具有很高的切割活性,其kcat值为031/min。
The ribozyme gene which specifically cleaved 12 lipoxygenase mRNA was designed,synthesized and cloned according to hammer head ribozyme model.A 25 bp long segment of 12 lipoxygenase mRNA was chemically synthesized as substrate,which was incubated with transcribed ribozyme RNA to detect its cleavage acitvity in vitro. The results showed that the ribozyme had a relatively high cleavage activity against 12 lipoxygenase mRNA segment at 37℃.It's k cat value and K m value was 0.083/min and 1300 nmol/L respectively.The ribozyme had a very high activity at 50℃ and its k cat value was 0.31/min.
出处
《生物工程学报》
CAS
CSCD
北大核心
2000年第1期96-98,共3页
Chinese Journal of Biotechnology
