马铃薯PGBSS—GUS融合基因在块茎中专一性表达的初步研究

从ＧＢＳＳ基因克隆上，酶切得到５上游区１．２ｋｂ的序列，与ＧＵＳ报告基基因融合，构建了ＰＧＢＳＳ１．２表达载体，通过农杆菌介导的方式，将ＰＧＢＳＳ１．２转和马铃薯品种Ｄｅｓｉｒｅｅ，卡那霉素筛选获得抗性再生植和微薯，利用ＰＣＲ特异性扩增和Ｓｏｕｔｈｅｒｎ Ｂｌｏｔｔｉｎｇ的方法证明了融合基因在马铃薯基因组中的整合，Ｘ－Ｇｌｕｅ活体染色表明，微薯切片具有高ＧＵＳ酶活性，而茎段中ＧＵＳ活性相对较低，初

A preliminary study on tuber-specific expression of P_(GBSS)-GUS fusion gene in transgenic potato

An expression vector pGBSS1. 2 was constructed by fusing 1. 2 kb 5' upstream region of GBSS(Granule-bound starch synthase ) gene with GUS (beta-glucuronidase ) reporter gene. After pGBSS1. 2 was introduced into Potato (Solantlln tubal-,)sunl c. v. Desiree ) plant by an Agrobacterium binary vector system. kanamycin-resistant plantlets and micro-tubers were obtained. X-Glue histochemical analysis showed high GUS activites in tubers,whereas these activities were very low in stems. By PCR and Southern blotting,it had been shown that this chimaeric pGBss-GUS gene was being integrated into genomic DNA of the regenerated transformants. Also,the 1. 2 kb promoter region of GBSS is highly-active and tuber-specific,and might be of great potential in potato bioreactor research.