木瓜輪點病毒鞘蛋白基因轉殖木瓜果實內鞘蛋白基因表現之探討

The Study of the Coat Protein Gene Expression in Fruit of PRSV CP-Transgenic Papaya

尤宗富(Tsung-Fu Yu) ； 林世敏(Shih-Min Lin) ； 鄧宇翔(Yu-Shang Teng) ； 溫銘嚞(Ming-Che Wen) ； 葉錫東(Shyi-Dong Yeh) ； 包慧俊(Huey-Jiunn Bau)

《臺灣農業化學與食品科學》 42卷6期 (2004/12) Pp. 466-473

https://doi.org/10.6578/TJACFS.2004.059

摘要

本研究目的在探討木瓜輪點病毒鞘蛋白基因轉殖木瓜果實中鞘蛋白轉基因之表現。以聚合酵素連鎖反應，可於成熟的基因轉殖木瓜果皮細胞中偵測到轉基因DNA的存在，但於果肉細胞則不易偵測到。在總蛋白質濃度比較上，基因轉殖木瓜與非基因轉殖木瓜並無顯著差異。在成熟果實上，以反轉錄－聚合酵素連鎖反應可偵測到果皮細胞中微弱的轉基因RNA存在，但以北方轉漬及西方轉漬法皆無法測得到轉基因 RNA 與其蛋白質於果皮及果肉細胞中的累積。另外，針對不同成熟度之基因轉殖木瓜果實中鞘蛋白轉基因之表現進行比較分析。結果以西方轉漬法無法測得到各不同成熟度之果肉中有轉基因蛋白質的累積，顯示基因轉殖木瓜中之鞘蛋白基因表現與其成熟度無關，且於細胞中之表現量極微。由以上結果顯示，食用基因轉殖木瓜後，由鞘蛋白所引起的致毒性及過敏原性之風險應極低。

關鍵字

木瓜輪點病毒 ； 鞘蛋白 ； 基因轉殖木瓜 ； 成熟度

並列摘要

The objective of this study was directed to analyze the expression and accumulation of the coat protein (CP) gene of Papaya ringsot virus (PRSV) in transgenic papaya fruit. Comparison of the level of total protein in, transgenic and non-transgenic fruits had no statistical difference. Using polymerase chain reaction (PCR) technique, the transgene was detected in the peels of transgenic fruits, but not in pulps. Transgene transcript was detected in transgenic fruit peels by reverse transcriptase-polymerase chain reaction, however, no positive signals were observed from transgenic fruit pulps. Results of northern blot analysis were similar to those of RT-PCR. Western blot analysis revealed that transgene protein accumulation was not detectable from transgenic fruit pulps at different maturity stages. Since the accumulation of transgene product, both at RNA and protein level, is not detectable by our methods. Our results indicated that the toxic and allergenic risks of taking the transgenic fruit are minimal.

並列關鍵字

Papaya ringspot virus (PRSV) ； Coat protein ； Transgenic papaya ； Maturity