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AcSERK1是菠萝(Ananas comosus)体细胞胚发生初期特异表达的基因,为探讨其转录调控规律,对其5′上游调控序列的转录起始位点及启动特性进行了鉴定。采用hiTAIL-PCR技术从菠萝基因组DNA中克隆了AcSERK1完整的5′上游调控序列,利用5′RACE技术鉴定出其转录起始位点位于起始密码子(ATG)上游258 nt(G)处。以AcSERK1 5′上游完整调控序列取代pBI121中的CaMV 35S启动子,构建植物表达载体AcSERK1(–2090/+258)︰︰GUS,并在菠萝不同组织器官中进行瞬时转化分析,发现AcSERK15′上游完整的调控序列(–2090/+258)能够启动GUS在胚性细胞中特异性表达,与AcSERK1表达规律相同,说明该启动子为胚性细胞特异性启动子,对调控外源基因在体细胞胚早期特异表达有重要意义。
AcSERK1 is an early specific expression gene of somatic embryos in Ananas comosus. In order to investigate its regulation of transcription, AcSERK1 was used to identify the transcriptional start site and start-up characteristics of the 5 ’upstream regulatory sequence. The full 5 ’upstream regulatory sequence of AcSERK1 was cloned from pineapple genomic DNA using hiTAIL-PCR technique. The 5’RACE was used to identify the transcription initiation site at 258 nt upstream of the start codon (ATG). The plant expression vector AcSERK1 (-2090 / + 258): ︰GUS was constructed by replacing the CaMV 35S promoter in pBI121 with the complete control sequence of AcSERK1 5 ’upstream and transient transformants in different tissues and organs of pineapple. It was found that AcSERK15’ (-2090 / + 258) can activate GUS specific expression in embryonic cells, which is the same as that of AcSERK1, indicating that the promoter is an embryogenic cell-specific promoter and plays an important role in regulating the expression of foreign genes in somatic embryos Early specific expression is of great significance.