关键词: 少根根霉, △6-脂肪酸脱氢酶基因, 油菜, γ-亚麻酸, 十八碳四烯酸

Abstract: 【Objective】 This study is to detect the expression of Rhizopus arrhizus Δ6-fatty acid desaturase gene RAD6 in transgenic rapeseed and provide the primary basis for the future application of gene engineering to GLA production. 【Method】 RAD6 was amplified and subcloned introduced expression vector pCPN2301 to generate a recombinant plasmid pCPNRAD6 under the control of CaMV35S promoter, which was subsequently transformed into Brassicu napus H165 by the Agrobacterium-mediated cotylendon node genetic transformation. The expression of target gene was detected by GC analysis. 【Result】 PCR, GUS staining, DNA dot and southern blot analysis of the transgenic plants showed that the target gene has been integrated into rape genome. Northern blot results proved that the target gene was expressed at mRNA level, and GC analysis indicated that RAD6 expressed in transgenic rapes with the accumulation ofγ-linolenic acid and octadecatetraenoic acid. However, with modification of sequences flanking AUG codon according to the Kozak sequence, a novel fragment (RAD6-1) of Rhizopus arrhizus Δ6-fatty acid desaturase gene was obtained and then transformed into Brassicu napus H165 for expression. As a result, compared with the transgenic rapeseed containing RAD6, the expression level ofΔ6-fatty acid desaturase gene was increased accordingly. 【Conclusion】 The expression system of Rhizopus arrhizus Δ6-fatty acid desaturase gene RAD6 in transgenic rapeseed was primarily constructed.

Key words: Rhizopus arrhizus, Δ6-fatty acid desaurase gene, Rapeseed, γ-linolenic acid, Octadecatetraenoic acid