Preliminary Study On Knockout Of GUX And ESK Genes In Rice Side Chain By CRISPR/Cas9 System

In recent years,hemi-fiber synthesis-related genes have become a hotspot.GUX and ESK affect the side chain synthesis of xylan,and the functions of GUX and ESK in plants are promising to further understand the synthesis of plant cell walls.In recent years,CRISPR / Cas9 system of plant genome fixed field of transformation has brought great changes in Arabidopsis,rice and other plants of endogenous genes targeted knockout application matures.In this study,GUO and ESK gene of rice rice were used as the research object,and their evolution,property and structure were analyzed by bioinformatics.The target knockout vector of multiple genes was constructed by optimizing the CRISPR / Cas9 system.Transformation and transgenic plants,lay a good foundation for the application of the system.The use of biological phylogenetic tree analysis GUX family gene.There are 3 GUXs in the aquatic plants of the lower plants,and there are three GUXs in the monocotyledonous rice.There are five GUXs in the Arabidopsis thaliana,10 in the eucalyptus,13 for the Eucalyptus,and 13 from the Eucalyptus In the process of evolution of higher plants to higher plants,GUX homologous genes increased from simp le to complex and diverse.The basic properties of GUX family genes in rice,gene structure analysis.GUX1 and GUX3 are transmembrane proteins.According to the transmembrane structure,the amino acid sequence in the transmembrane region is 1 ~ 52,1 ~ 27,while GUX2 does not exist in the transmembrane domain.In the GUX gene family amino acid analysis,GUX1,GUX2,GUX3 between the differences.The use of biological phylogenetic tree analysis ESK family gene.Among them,there are one of the lower aquatic plants,three in the rice,six in the Arabidopsis,15 in the poplar,13 in the Eucalyptus,and from the lower plants to the higher plants,ESK homologous genes increase from simple to complex and diverse trends.According to the analysis of ESK1 ~ 3 amino acid sequence,ESK2 is a transmembrane protein.According to the transmembrane structure,the amino acid sequence in the transmembrane region is 1 ~ 20,while ESK1 and ESK3 are absent.In ESK2 gene family amino acid analysis,ESK1,ESK2,ESK3 differences between.The expression levels in different tissues were analyzed by fluorescence quantitative PCR.Expression of different tissues: stem> root> leaf> seedling.Using the CRISPR / Cas9 system to construct rice three target carriers.By using the conventional PC R detection of GUX transgenic rice,11 rice positive seedlings were obtained by the optimization of the Cas9 protein vector.The positive rate was 100%.ESK transgenic rice routine PC R detection,obtained 2 strains.The transgenic plants obtained are not a lot.