大豆异黄酮合酶基因的克隆及序列分析

目的:大豆异黄酮是多酚类混合物,有防治肿瘤发生,提高机体免疫力等多种保健功能。异黄酮合酶(isoflavone synthase,IFS)是合成异黄酮的关键酶。本文为了利用异黄酮的特有生物学功能,从大豆中克隆了该基因。方法:采用PCR扩增从大豆Glycine max(Linn.)Merr.]总RNA中分离了异黄酮合酶基因,并将其克隆到pUCm-T载体并测序。结果:得到全长1583bp的片段。以期用于构建诱导表达基因敲除系统,并用于无性繁殖植物的无标记基因转化。结论:序列分析表明,异黄酮合酶基因(IFS1)含1583个核苷酸,与已报道的序列比较,核苷酸的同源性为92%。

Cloning and Sequence Analysis of Isoflavone Synthase from Soybean

Objective: The soybean isoflavone,a polyphenol mixture,benefits our health care such as tumor prevention and cure,immunity raise.Isoflavone synthase(IFS) is a key enzyme in the synthesis of isoflavone.For making good use of the specific biological function of isoflavone,A cDNA fragment from full length IFS gene was amplified from Glycine max(Linn.) Merr.Methods: A cDNA fragment from full length IFS gene was amplified by polymerase chain reaction from Glycine max(Linn.) Merr.,and it was sequenced after cloned into pUCm-T vectors.Results: A cDNA fragment of 1 583bp from full length IFS gene was got.They constructed the induced gene knock out system,to use into markers free transgenic plant. Conclusion: The sequencing results indicated that the cloned fragment of ifs1 contained 1 583 nucleotides,and shared a sequence homology of 92% with that from Genbank accession number AF195798(IFS1).