CAJ | 학술논문

通过RT-PCR,结合RACE技术,得到了玉米(Zea mays L.)果糖-6-磷酸,2-激酶/果糖-2,6-二磷酸酶的全长cDNA克隆,命名为mF2KP.氨基酸序列同源性比较发现,mF2KP蛋白可以分为两个部分:C端包含高度保守的催化功能区,N端为植物中特有的多肽.将mF2KP基因中一段包含完整催化功能区的片段在大肠杆菌(Escherichia coli)中表达,融合蛋白具有果糖-6-磷酸,2-激酶/果糖-2,6-二磷酸酶活性.Northern杂交证明在种子活力不同的幼苗中,mF2KP的转录水平存在明显差异.种子活力越高,幼苗中mF2KP的转录水平越低.
통과RT-PCR,결합RACE기술,득도료옥미(Zea mays L.)과당-6-린산,2-격매/과당-2,6-이린산매적전장cDNA극륭,명명위mF2KP.안기산서렬동원성비교발현,mF2KP단백가이분위량개부분:C단포함고도보수적최화공능구,N단위식물중특유적다태.장mF2KP기인중일단포함완정최화공능구적편단재대장간균(Escherichia coli)중표체,융합단백구유과당-6-린산,2-격매/과당-2,6-이린산매활성.Northern잡교증명재충자활력불동적유묘중,mF2KP적전록수평존재명현차이.충자활력월고,유묘중mF2KP적전록수평월저.
A full-length cDNA encoding fructose-6-phosphate, 2-kinase/fructose-2,6-bisphosphatase from maize (Zea mays L.) was cloned by the methods of reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE), and designated as mF2KP. The encoded protein is composed of two regions. Its COOH-terminal region is catalytic region and homologous to the enzymes from other eukaryotes; and its NH2-terminal region is common and special region only in plant. A truncated fragment of mF2KP covering integrated catalytic region was expressed in Escherichia coli. The fusion protein had the activities of fructose-6-phosphate, 2-kinase as well as fructose-2,6-bisphosphatase. Northern blot showed that the transcript level of mF2KP in seedlings initiated from strong-vigor seeds is lower than that from weak-vigor seeds.