摘要
将０．４、０．８、１．６、２．９ｋｂＧＢＳＳ基因的５′侧翼区与ＧＵＳ基因融合，构建了双元表达载体。０．８ｋｂＧＢＳＳＧＵＳ通过基因枪介导在块茎切片中获得了瞬间表达。以上建构物通过农杆菌介导转入了马铃薯（ＳｏｌａｎｕｍｔｕｂｅｒｏｓｕｍＬ．ｃｖ．Ｄｅｓｉｒｅｅ）。ＸＧｌｕｃ染色及ＰＣＲ结果证实已获得转基因植株。利用离体块茎诱导系统，ＧＵＳ表达用荧光进行定量检测，结果显示，２．９、１．６、０．４ｋｂＧＢＳＳＧＵＳ的表达均以块茎明显高于茎段，达２～１０倍。０．８、１．６、２．９ｋｂＧＢＳＳＧＵＳ表达高于０．４ｋｂＧＢＳＳＧＵＳ。蔗糖浓度的升高可诱导ＧＢＳＳＧＵＳ的表达，而光照抑制了ＧＢＳＳＧＵＳ的表达。
Binary vectors were constructed by fusing 0.4, 0.8, 1.6, and 2.9 kb 5′ flanking regions of GBSS gene with GUS (β glucuronidase). Transient GUS expression was observed in in vitro tuber slices bombarded with 0.8 kb GBSS GUS construct. These constructs were then transferred into potato ( Solanum tuberosum L. cv. Desiree) via Agrobacterium tumefaciens transformation. Transgenic potato plants were confirmed by X Gluc staining and PCR. Using in vitro tuberization system, GUS expressions were assayed with fluorescence, it was shown that 0.8, 1.6 and 2.9 kb GBSS GUS expressions were higher than 0.4 kb GBSS GUS. 1.6 and 2.9 kb GBSS GUS expressions were about 2 to 10 folds higher in tubers than in stems. In cultured shoots, GBSS GUS expression could be induced by increased sucrose concentration but inhibited by light.
关键词
马铃薯
基因调控
GBSS基因
块茎专一表达
诱导
Gene regulation, Granule bound starch synthase, Tuber specific expression, in vitro tuberization, Solanum tuberosum
