β-1,3-葡聚糖酶基因植物表达载体的构建

CONSTRUCTION OF PLANT EXPRESSION VECTORSHARBORING 1,3GLUCANASE GENE FROM BARLEY

采用ＰＣＲ的方法，对来自大麦ｃＤＮＡ克隆的β－１，３－葡聚糖酶基因（β－１，３－ｇｌｕｃａｎａｓｅ）进行了扩增，在此基因的５’及３’端分别加入了克隆所需的ＸｂａⅠ和ＳｓｔⅠ限制性酶切位点，定向克隆到经改造的质粒载体ｐＢｉｎｈ中，获得了含有β－１，３－葡聚糖酶基因的植物表达载体ｐＢｉｎｈ－Ｇｌｕ。该载体含有ＣａＭＶ３５Ｓ启动子、ＮＯＳ终止子及具有卡那抗性的ＮＰＴⅡ基因。

glucanase cDNA clone from barley has been amplified by PCR method. Through primers design a Xba site and a Sst site have been added to 5end and 3end of the gene respectively. A plant expression vector pBinhGlu was constructed by inserting modified 1,3glucanase gene into modified plasmid pBinh vector. There are CaMV 35S promoter, NOS terminator and Kan R NPT gene elements in this vector.