Investigation On The Effect Of Ribosomal Protein L36 Mutation On Cell Growth In Saccharomyces Cerevisiae

Ribosome synthesis is a highly complex and coordinated process. Variousribosomal proteins have been reported to be involved in pre-rRNA processing,ribosomal subunits assembly, and ribosomal functions pertaining to translation.However, the detailed mechanisms how these different ribosomal proteins contributeto the above-mentioned reactions are still not fully understood. Recent study on yeastribosomal protein RPS 14 has set a model to investigate the functions of ribosomalproteins on ribosome biogenesis. In this work, we have constructed a yeast strain inwhich the two endogenous RPL36 genes, RPL36A and RPL36B, have been deletedby homologous recombination. This RPL36 genes deleted strain can survive and bemaintained due to the presence of a transformed wild type RPL36B gene constructedin a plasmid containing Ura selection marker. Through plasmid shufflingmanipulation, seven mutated RPL36B constructs were introduced into the deletedstrain to replace the wild type RPL36B gene in the Ura plasmid as the only source ofRPL36 protein. We have observed that some mutations can clearly affect the growthrate of yeast. This collection of mutant strains will be used to investigate the functionsof RPL36 proteins in pre-rRNA processing, maturation of ribosomal subunits andfunctions of assembled ribosomes. The results gathered from this study will help usunderstand more about the roles of RPL36 in eukaryotic ribosomes.