作者: SV.Wesley (2001)
载体名称: pHELLSGATE12
质粒类型: 基因沉默载体;Gateway载体;植物双元表达载体
高拷贝/低拷贝: 高拷贝
克隆方法: Gateway
启动子: CaMV 35S
载体大小: 17861 bp
5' 测序引物及序列: 35S promoter:5’CTATCCTTCGCAAGACCCTTC 3’
3' 测序引物及序列: --
载体抗性: 壮观霉素和链霉素
筛选标记: 卡那霉素
克隆菌株: DB3.1
宿主细胞(系): 植物细胞、农杆菌
备注: pHELLSGATE12载体是高效、高通量、有效的植物基因沉默载体。
产品目录号: --
稳定性: 稳表达
组成型/诱导型: 组成型
病毒/非病毒: 非病毒





Post-transcriptional silencing of plant genes using anti-sense or co-suppression constructs usually results in only a modest proportion of silenced individuals. Recent work has demonstrated the potential for constructs encoding self-complementary 'hairpin' RNA (hpRNA) to efficiently silence genes. In this study we examine design rules for efficient gene silencing, in terms of both the proportion of independent transgenic plants showing silencing, and the degree of silencing. Using hpRNA constructs containing sense/anti-sense arms ranging from 98 to 853 nt gave efficient silencing in a wide range of plant species, and inclusion of an intron in these constructs had a consistently enhancing effect. Intron-containing constructs (ihpRNA) generally gave 90-100% of independent transgenic plants showing silencing. The degree of silencing with these constructs was much greater than that obtained using either co-suppression or anti-sense constructs. We have made a generic vector, pHANNIBAL, that allows a simple, single PCR product from a gene of interest to be easily converted into a highly effective ihpRNA silencing construct. We have also created a high-throughput vector, pHELLSGATE, that should facilitate the cloning of gene libraries or large numbers of defined genes, such as those in EST collections, using an in vitro recombinase system. This system may facilitate the large-scale determination and discovery of plant gene functions in the same way as RNAi is being used to examine gene function in Caenorhabditis elegans.




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