Nalysis Of CDNA Microarray For Rice Roots Under Fe-Deficiency Stress And Molecular Cloning Of CSN6 Gene

Iron deficiency in plants is a widely-existed problem mainly for the sake of agricultural meaning , and then causing a large amount of scientists engage into this field . Among them , the studies around rice plant is of paramount important because of its wide usage both as food in agriculture and as a model plant in professional research works. People try to uncover the myth of the iron uptake mechanism in rice so we can get some basic understandings about the iron metabolism in plants.In this work , we implied cDNA microarray technology screening the differential expressed genes between 5 days - iron deficiency- and iron sufficiency- treated rice root. Totally , there were 451 genes detected in our microarray experiment , among which there were 203 genes presented up-regulated expression mode in the iron deficiency samples , and 248 genes down-regulated. these 451 genes can be divided into several categories, i.e. plasma membrane and transporter genes ; cytoskeleton genes ; vesicle transportation genes ; material and energy metabolism genes ; regulation factor genes ; cell cycle control genes ; cell signaling genes , etc . using BLASTn and PubMed in NCBI we get the detail information about these genes , described there molecular functions , and suggested the roles they may play in iron deficiency tolerance.We chose the COP9 signalosome subunit6 gene present in 451 genes described above for molecular cloning work. We got 990bp fragment containing the full length of the CSN6 cDNA through RT-PCR , integrated it with the pMD-18-T cloning vector . For further research work of cellular localization and functional analysis , we combined the CSN6 cDNA with the pCAMBIA 1302 expression vector used specially in plant gene engineering , got cis - and trans- recombinant . finally , we implied the RealTime PCR technology to test the expression mode of CSN6 Cullin CDC21 and permease gene in 7 day-long period .Results showed some unparallel to the microarray datum , but in all, we concerned these results were of coincidence , which suggested that these genes interplayed each other during the iron deficiency period.