Some Fission Yeast Ribosomes Genes And Gene Knockout And Phenotypic Comparative Study

Ribosomal protein (ribosomal protein, RP) is the important composition of ribosomes unit, its main function is to participate in the assembly and protein ribosomes translation. Ribosomes are completely assembled by a large ribosomes protein and ribosomes RNA (rRNA) in nucleoli.There are about150species of rRNA genes and137species of ribosomal pro-tein coding genes as a candidate gene in the great project of transcrip-tion. The lab early has successfully knocked out ribosomal protein gene L32-1and L32-2, and found that the knockout strains in liquid medium (YPD), appear hundreds of cells agglutinate together forming visible groups as a floe phenomenon (flocculation). In order to understand the cause of yeast cells flocculatIon.this article on this basis, the random selection of knockout ribosomal protein gene L21-2and L9-2.The same time, we also knockout the genes with don’t participate in the ribosomes translation,called SPBC1347.09, SCK1, SPAC732.02c, SPBC13G1.02. By PCR, we obtained upstream and downstream genes and screening of resistance gene KanMX6of knockout fragment. Finally three fragments joined in the PCR tube together, with the upstream and downstream homologous arm of P1and P2to PCR the full-length knockout fragment. Application of Li-Ac transformation method, the knockout segments through into recipient strain SP-Q01, YPD liquid medium trained and G418resistance plate to screen positive clones. With the extracting genomic of positive clones,PCR to verify which the correct positive strains preservation.Through the analysis on these six knockout strains of the training, cells form of observation and the determination of the growth curve,we found that ribosome protein gene knockout strains grew slowly, and had flocc-ulation; And unless ribosome protein gene was knocked, or disabled,no this phenomenon. To this we can crack prelimin-ary judgment about bud ding yeast cells that ribosomal proteins express inadequate can cause yeast cells flocculation. For the specific flocculation mechanism, we also need to further validation.