Construction Of PSupper1300-EB1c-GFP Recombinant Plasmid And The Screening Of Transgenic Plants

All eukaryotes must pass parental inherited message to offspring through cell abruption. And the reorganization of the microtubules have a hand in this procedure. There is an increase in MT dynamics which occurs concomitantly with nuclear membrane disruption that could be important for microtubules morphogenesis.The dynamics of microtubules are not only related to their intrinsic dynamic instability but also monitored by microtubule-associated protein(MAPs). MT plus-end-tracking proteins(+TIPs) that is a category of proteins of MAPs can recognize the distal part of a polymerizing MT. +TIPs are the regulation factors of microtubule dynamics, End-binding protein 1(EB1) is the core of the microtubule plus-end complex, and is the main regulation factor of microtubule dynamics. EB1 is necessary for karyokinesis fusiform body microtubule location and chromosome abruption. Loss of EB1 function causes defects in karyokinesis fusiform body microtubule structure in metaphase cells. Whether chromosome segregation will be abnormal under the circumstances is still not clear, and what about it to induce chromosome segregation by regulating spindle movement is still not fully clear. Han et al. found that abnormal abruption would be happened in root-tip cells of wheat seedlings under the enhanced UV-B eradiation, and the chromosomes wouldbe three, four and six bundles. Whether EB1 has a function under the circumstances require to further be tested. Meanwhile, whether the strengthened UV-B eradiation can cause the change of microtubule distribution in plant root cells and EB1 has an significant function under the circumstances remains to be further studied.In this study, primers are devised according to the order delivered in Gen Bank, the young leaves of wild type Arabidopsis thaliana were used, first, overall RNA was picked up and then EB1 c was amplified. Finally, the p Supper1300-EB1c-GFP expression vector was builded after p UC19-EB1 c cloning vector was constructed and transfected Arabidopsis thaliana plants,transgenic plants were filtered. At last, p UC19-EB1 c cloning vector and recombinant plasmid p Supper1300-EB1c-GFP had been got.After sequencing, the order of the EB1 c were contrasted against the delivered order in Genbank, and the homology was 100%.Screening the transgenic Arabidopsis seeds after the agrobacterium infected, and the T2 progeny were acquired.