Isolation and Characterization of Functional Ribosomal RNA Fragments

;​R o o t E n t y F C o m p o b j b W o r d d o c u m e n t O b j e c t P O O l / Q /问1 2 3 4 5 6 7 8 9：;F Microsoft Word 6.0文档MSWordDoc。6;​V t C；r 9^ u F ^ ~ t ~ u F v j v F)F ~ tT ~ tNj；j V P ' ' V G # V F j V F P O ~ u V V Pj @ # 3 V Vj V ' Vj j V V V Pj Vj 2# ~ V ！本研究的重点是核糖体RNA不同区域的功能。提出了一种识别功能重要rRNA片段的新方法。也就是说，随机rRNA片段的表达文库将筛选能够影响体内核糖体功能的RNA物种。该方法的有效性已在初步研究中得到验证，从随机的rRNA片段文库中分离出对核糖体靶向抗生素红霉素具有抗性的活性rRNA片段。分析红霉素耐药RNA的功能是研究的目的之一。此外，功能选择方案将用于鉴定表达其他活性rRNA片段的克隆，这些片段1)改变对核糖体靶向抗生素的敏感性，2)干扰核糖体组装和rRNA加工，3)影响翻译的准确性，4)干扰蛋白质生物合成。遗传和生化分析将用于确定所选rRNA片段生物活性的结构要求及其作用模式。这将有助于更好地理解核糖体中相应rRNA区域的功能。细胞中的所有蛋白质都是由核糖体合成的。核糖体由核糖体RNA和蛋白质分子组成。核糖体RNA的功能是本次研究的主题。拟议的研究将采用一种新方法。核糖体RNA将被随机切割成小块。将分离功能片段并对其功能进行研究。将尝试鉴定能够影响核糖体组装、蛋白质合成准确性和对抗生素敏感性的核糖体RNA片段。总的来说，所提出的方法应该有助于更好地理解rRNA在蛋白质生物合成过程中的功能。*** @ ....()()))()() y S u m m r I f n o r m t I o n ( 0Oh +' 0 $ H l D H R:\WWUSER\TEMPLATE\NORMAL。marcia steinberg @ C @ @ C @ Microsoft Word 6.0 2;3 y C 0 ！T；本研究的重点是核糖体RNA不同区域的功能。提出了一种识别功能重要rRNA片段的新方法。也就是说，随机rRNA片段的表达文库将筛选能够影响体内核糖体功能的RNA物种。该方法的有效性已在初步研究中得到验证，从随机的rRNA片段文库中分离出对核糖体靶向抗生素红霉素具有抗性的活性rRNA片段。功能分析

; R o o t E n t r y F C o m p O b j b W o r d D o c u m e n t O b j e c t P o o l /Q /Q 1 2 3 4 5 6 7 8 9 : ; F Microsoft Word 6.0 Document MSWordDoc Word.Document.6 ; V t C; r 9^ u F ^ ~ t ~ u F v j v F )F ~ tT ~ tNj;j V P " V P G # V F j v F P O ~ u hd V Pj @ # 3 v v j V ' v j j v V Pj Vj 2# ~ v! v N )F ~ v Wh $ 'Nu v v v $9420768 Mankin The focus of this research is the function of different regions of ribosomal RNA. A new approach for identification of functionally significant rRNA segments is proposed. Namely, an expression library of random rRNA fragments will be screened for RNA species which can affect ribosome functions in vivo. The efficiency of the approach has been tested in preliminary studies where an active rRNA fragment conferring resistance to the ribosome targeted antibiotic erythromycin has been isolated from a random library of rRNA fragments. Analysis of the function of the erythromycin resistance RNA is one of the aims of the research. Further, functional selection protocols will be used to identify clones which express other active rRNA fragments that 1) alter sensitivity to ribosome targeted antibiotics, 2) interfere with ribosome assembly and rRNA processing, 3) affect accuracy of translation and 4) interfere with protein biosynthesis. Genetic and biochemical analysis will be used to determine structural requirements for biological activity of the selected rRNA fragments and the mode of their a ction. This should lead to a better understanding of the functions of corresponding rRNA regions in the ribosome. %%% All proteins in the cell are synthesized by ribosomes. The ribosome is composed of ribosomal RNA and protein molecules. Function of the ribosomal RNA is the subject of the proposed research. A new approach will be used in the proposed research. Ribosomal RNA will be cut into small random pieces. Functional fragments will be isolated and their function studied. An attempt will be made to identify fragments of the ribosomal RNA that can influence assembly of the ribosome, accuracy of protein synthesis and sensitivity to antibiotics. In general, the proposed approach should lead to a better understanding of the functions of rRNA in the process of protein biosynthesis. *** @ ....()()))()() S u m m a r y I n f o r m a t i o n ( 0 Oh +' 0 $ H l D h R:\WWUSER\TEMPLATE\NORMAL.DOT 9420768 marcia steinberg marcia steinberg @ C @ @ C @ Microsoft Word 6.0 2 ; e 3 e y C 0 ! T u ; h 9420768 Mankin The focus of this research is the function of different regions of ribosomal RNA. A new approach for identification of functionally significant rRNA segments is proposed. Namely, an expression library of random rRNA fragments will be screened for RNA species which can affect ribosome functions in vivo. The efficiency of the approach has been tested in preliminary studies where an active rRNA fragment conferring resistance to the ribosome targeted antibiotic erythromycin has been isolated from a random library of rRNA fragments. Analysis of the functio