拟南芥CRY1基因C末端导入甘蓝型油菜的遗传及表达

Inheritance and expression of the C-terminal fragment of Arabidopsis Cryl gene transferred into Brassca. napus L.

通过卡那霉素抗性鉴定,对甘蓝型油菜westar转基因后代T2代株系进行了研究。结合PCR检测及序列比对验证,对拟南芥CRY1基因C末端导入甘蓝型油菜后的遗传表现进行了分析,并利用GUS染色和花色素苷积累量的比较,对CRY1基因C末端在转基因油菜中的表达进行了研究。结果表明：转化的外源目的基因多数以嵌合形式,少数以单拷贝和嵌合的混合形式整合到转基因油菜的基因组中;目的基因能稳定地遗传给后代,株系间遗传传递率为62.5%;T2代的遗传行为少数呈现孟德尔遗传,多数呈现非孟德尔遗传现象;拟南芥CRY1基因能在转基因油菜中表达,但也出现了沉默现象。

The T2 generation of the transgenic T1 progeny in B. napus L. cv. westar, which was obtained by Floral-dip method, was studied with Karamicine（Km） test and the inheritance of the C-terminal fragment of Arabidopsis CRY1 gene was analysed. The positive plants from Km test were examinated with PCR detection and the specific DNA fragment was isolated, sequenced and compared to the original target gene sequence. GUS examination and anthoeyanin accumulation in the PCR-positive plants were further examinated to study the experssion of the target gene in the transgenic plants. Theresults indicated that the C-terminal fragment of Arabidopsis CRY1 gene was successfully integrated into the rape genome, mainly in the form of tabling, with a few in the form of mixed copy of single and tabling. The transformed gene could be stably transmitted in the progeny, with a transmitted line rate of 62.5 %. The genetic pattern of the transformed gene in the T2 generation was mostly non-Mendelian. The transgene was expressed in some of the tmnsgenic plants, but kept silent in other plants.