木薯AGPase基因的克隆与载体构建

Cloning and Expression Vector Construction of AGPase Gene from Cassava

为了研究木薯淀粉合成酶关键基因AGPase在木薯淀粉合成中的作用,根据GenBank中其他物种AGPase小亚基基因序列,从高淀粉木薯‘辐选01’中克隆得到含3’末端的909bp的基因片段,回收克隆测序。结果显示,克隆获得片段与基因库中登录的蓖麻、毛果杨、甜橙、番薯、蜜柑、大豆同源性依次达到91%、91%、87%、86%、86%及85%。将阳性克隆与PBI121载体进行双酶切、连接,构建植物反义表达载体PBI121-AGPss,并将其导入农杆菌,得到农杆菌工程菌株。为进一步开展木薯淀粉合成相关的基因调控、转基因及基因功能研究奠定基础。

In order to study the role of the key enzyme gene AGPase in starch synthesis,according to the other species’AGPase small subunit gene of the GenBank,we got 909 bp gene fragment with 3’end from FX01.Through sequence similarity search results showed that,its nucleotide sequence with GenBank landing Ricinus communis,Populus trichocarpa,Citrus sinensis,Ipomoea batatas,Citrus unshiu and Glycine max sequence homologily,respectively of 91%,91%,87%,86%,86% and 85%.AGPase gene antisense expression vector PBI121-AGPss was constructed based on the known vector pBI121,and transferred it into Agrobaterium tumefaciens to get the transformed bacteria.It made the foundation for further study of cassava starch synthesis-related gene regulation,GM and gene function.