根癌农杆菌介导的北极悬钩子（Rubus arcticus L.）转化

Transformation of arctic bramble (Rubus arcticus L.) by Agrobacterium tumefaciens.

作者信息

Kokko H I, Kärenlampi S O

机构信息

Department of Biochemistry and Biotechnology, University of Kuopio, P.O. Box 1627, FIN-70211 Kuopio, Finland Fax: +358-71-2811510 e-mail:

出版信息

Plant Cell Rep. 1998 Jul;17(10):822-826. doi: 10.1007/s002990050491.

DOI:10.1007/s002990050491

PMID:30736600

Abstract

Genetic transformation of arctic bramble (Rubus arcticus L.) was achieved utilizing a Ti-plasmid vector system of Agrobacterium tumefaciens. Internodal stem segments were inoculated with Agrobacterium strain EHA101 carrying a T-DNA with the CaMV 35 S promoter-gus-int marker gene from which β-glucuronidase (GUS) is expressed only in plants. Regenerants were produced on Murashige and Skoog medium. Growth of Agrobacterium was inhibited with cefotaxime. Kanamycin was used as the selective agent for the transformants. Regenerants were assayed by histochemical GUS staining, and by Southern analysis using a gus-int probe. Transgenic arctic bramble plants containing gus-int and expressing GUS were recovered. Expression has been stable for 3 years in micropropagation.

摘要

利用根癌农杆菌的Ti质粒载体系统实现了北极悬钩子（Rubus arcticus L.）的遗传转化。用携带带有CaMV 35S启动子 - gus - int标记基因的T - DNA的农杆菌菌株EHA101接种节间茎段，其中β - 葡萄糖醛酸酶（GUS）仅在植物中表达。在Murashige和Skoog培养基上产生再生植株。用头孢噻肟抑制农杆菌的生长。卡那霉素用作转化体的选择剂。通过组织化学GUS染色和使用gus - int探针的Southern分析对再生植株进行检测。回收了含有gus - int并表达GUS的转基因北极悬钩子植株。在微繁殖中，表达已稳定3年。