Validation of cDNA Microarray as a Prototype for Throughput Detection of GMOs

為了發展出一套有效且快速的基因改良食品檢驗系統，本實驗搜集了一般常見於基因改良作物的外來DNA基因片段如花椰菜鑲嵌病毒CaMV 35S啟動子，NOSt或35S終結子、nptⅡ、hph或pat篩選基因、gus或gfp標示基因，cp4 epsps、cryl Ab等己知的轉殖基因和4個植物的內標準基因（如invertase, legumin, β-tubulin, actin基因）等，打點於微型晶片上建立出一個大量篩檢基因改良食品的cDNA微矩陣系統。研究中先以本實驗室自行完成的轉殖水稻、蕃茄及馬鈴薯做初步的檢測和系統敏感度測試。結果顯示，外來基因CaMV 35S啟動子、NOSt、35S終結子、nptⅡ和hph等基因都能成功地檢測出來，而相對應的野生種則只檢測出4個植物的內標準基因。為了更進一步了解此系統對市售樣品的檢測效率，實驗中也測定GM馬鈴薯及GM玉米及玉米脆片食品和市售小麥種子，結果同樣可以明確的偵測到外來DNA基因的存在。而在以馬鈴薯所做的敏感度測試中也顯示此系統至少可以偵測到原料中只含有0.01-0.002%的GM馬鈴薯塊莖，因此證明這個cDNA微矩陣系統可以發展成一個快速且便於大量篩檢的檢測系統。

關鍵字

cDNA微矩陣法；基因轉殖物種GMO ；基因轉殖馬鈴薯；基因轉殖玉米

並列摘要

As a step towards developing an efficient genetically modified organism (GMO) detection system, the present investigation proposes microarray as a prototype for high-throughput detection of pure GMO samples. Common T-DNA regions of the expression cassette such as 35S promoter, 35S terminator, nopaline synthase terminator (NOSt), hygromycin, and kanamycin selection marker genes were detected in the transgenic rice, tomato, and potato developed in our laboratory. For proof-of-concept purposes, marketintroduced GM potato, GM maize, and cornflakes were screened. The non-GM potato and maize control detected only the presence of endogenous genes while the GM targets detected the presence of transgenic genes such as CaMV 35S promoter, 35S terminator, NOSt and nptⅡ, pat, cp4 epsps, cryl Ab genes on chips. Moreover, it was observed that the sensitivity of this system for serially diluted GM potato tubers was up to 0.01-0.002% of the mass fraction. Due to its high accuracy and speed, it is believed that the microarray detection system will play an important role in routine, high-throughput detection of pure GMO samples in the future.

並列關鍵字

cDNA microarray ； Genetically modified organisms GMO ； GM potato ； GM maize

國際替代計量

Validation of cDNA Microarray as a Prototype for Throughput Detection of GMOs

全文下載

主題瀏覽