SEQUENCING OF THE U6 PROMOTERS IN CASTOR BEANS AND VECTOR CONSTRUCTION FOR CRISPR/Cas9 GENOMIC EDITING ON THEIR BASIS

Castor bean is an important crop in many countries. It mainly used to obtain the castor oil, which widely applied various industries. The rich protein cakes and meals are remains after oil pressed. They promising for use as additives fodder production. However, it limited due presence of toxins. These ricin ricinine alkaloid. One such methods can be genomic editing using CRISPR/Cas9 system. technology consists cutting target region intact DNA by Cas9 enzyme with assistance a short guide RNA fragment. delivery genes into cell edited plant often carried out plasmid vectors. For efficient synthesis vectors, promoters small nuclear usually used. In dicotyledonous plants editing, promoter Arabidopsis U6 gene most this work, amplification, sequencing analysis were at first time. obtained sequences analyzed construction vectors editing. Our aim was study help bioinformatics molecular genetic approaches. Zanzibar Green Gibzonskaya varieties work. isolated from young leaves. preparation sequences, alignments homology level calculation GenDoc program (http://www.nrbsc.org/gfx/genedoc/index.html). Primers amplification designed Primer3 (http://bioinfo.ut.ee/primer3-0.4.0/). PCR performed C-100 machine (Bio-Rad Laboratories, Inc., USA). products separated 1.5 % agarose gel 6 B/cm Sub-Cell GT electrophoresis camera amplicons purified GeneJET Purification Kit (Thermo Fisher Scientific, cloned pAL2-T vector (Evrogen, Russia) according protocol manufacturer. construction, pRGE31, HindIII SbfI restriction enzymes (Sibenzyme, nucleotides digested Gel Extraction Bioinformatic search conducted GenBank base, 12 scaffolds found. Six USE TATA-box elements regulation primer design cv. matrices. fragment sequenced. amplicon reveled that regions two studied similar. identity different ranged within 51-77 %. comparison these ones other plants, 42-64 motifs involved synthesis.