CAJ | 학술논문

利用RT-PCR和RACE技术从白桦(Betula platyphylla)中克隆了编码苯丙氨酸解氨酶(PAL)的cDNA,其2322bp的ORF编码773个氨基酸,其推导的氨基酸序列包含PAL-HAL和PAL2个功能域以及酶活性中心序列GTITASGDLVPLSYIA,该序列同其它5种植物的序列一致性为60%～73%,其中与美洲红(Rhizophora mangle)树最高为73.1%.以该序列构建了系统进化树,白桦与美洲红树聚为一类,其余3种裸子植物长白松、沙地海岸松和银杏聚为一类.BplPAL1基因在各组织中均有不同的转录表达,在次生木质部表达最强,其次是幼叶,在花序中的表达量较低,说明BplPAL1基因在各组织中的调控和表达是不同的.
이용RT-PCR화RACE기술종백화(Betula platyphylla)중극륭료편마분병안산해안매(PAL)적cDNA,기2322bp적ORF편마773개안기산,기추도적안기산서렬포함PAL-HAL화PAL2개공능역이급매활성중심서렬GTITASGDLVPLSYIA,해서렬동기타5충식물적서렬일치성위60%～73%,기중여미주홍(Rhizophora mangle)수최고위73.1%.이해서렬구건료계통진화수,백화여미주홍수취위일류,기여3충라자식물장백송、사지해안송화은행취위일류.BplPAL1기인재각조직중균유불동적전록표체,재차생목질부표체최강,기차시유협,재화서중적표체량교저,설명BplPAL1기인재각조직중적조공화표체시불동적.
The cDNA (BplPAL1) encoding phenylalanine ammonia-lyase (PAL) were cloned from Betula platyphylla by reverse transcription polymerase chain reaction (RT-PCR) and 5' and 3' rapid amplification of cDNA ends (RACE), which contains an open reading frame (ORF) (2322bp) encoding 773 amino acids. The amino acid sequences contain two functional domains of PAL-HAL and PAL, and the sequence of enzyme active site (GTITASGDLVPLSYIA), which has 60%～73% of consistency with other five plant species and the consistency with Rhizophora mangle is the highest (73.1%). Phylogenetic analysis of BplPAL1 with other plant species revealed that B. platyphylla and R. mangle formed a gymnosperm-type PAL subfamily, and Pinus sylvestris, Pinus pinaster, Ginkgo biloba formed a subfamily. Semi-quantitative RT-PCR analysis indicated that the expression of BplPAL1 genes was different in tissues, exhibiting the strongest in xylem, next in young leaves, and the weakest in inflorescences. The results suggest that BplPAL1 genes are differently regulated and expressed in tissues.