To investigate biological roles of Myosin class I in the morphology of M. circinelloides and identify good candidates for further analyses of Mucor pathogenesis, we are going to inhibit their expression using RNAi technique. We named the two myosin class I like genes as M1L1 (ID 140500) and M1L2 (ID 116620). In the present study, we report the results of generation of RNAi plasmid to inhibit the expression of M1L1 gene. | HNUE JOURNAL OF SCIENCE Natural Sciences 2020 Volume 65 Issue 4A pp. 27-32 This paper is available online at http GENERATION OF RNAi PLASMID CORRESPONDING TO THE m1l1 GENE ID 140500 BELONGING TO MYOSIN CLASS I IN THE FUNGUS Mucor circinelloides Le Ngoc Mai1 Le Phuong Thao1 Pham Thi Thu Trang 2 and Trieu Anh Trung1 1 Faculty of Biology Hanoi National University of Education 2 Nguyen Trai Gifted High School Hai Duong province Abstract. Mucormycosis has emerged as a second most common filamentous fungal infection in human caused by species belonging to order Mucorales especially in immunocompromised patients with a high mortality ratio. Our understanding of its virulence determinants is limited leading to the lacking of efficient therapies. Dimorphism is one of the phenotypic characteristics that can be used as a marker for virulence screening. Mucor circinelloides a dimorphic fungus has become an attractive model for studies on various studies including mucormycosis. Our previous study identified Mcmyo5 protein belonging to Myosin class V which plays an important role in Mucor pathogenesis. We have found three protein families with high similarities of amino acid sequences compared to Mcmyo5 including Myosin class I II and V in the Mucor genome. To investigate the possible roles of those genes in Mucor pathogenesis using RNAi techniques we generated the RNAi plasmid which is corresponding to endogenous gene m1l1 ID 140500 . In the present study we show the results of isolation and cloning a fragment of the target gene m1l1 into the vector pMAT1812 to obtain RNAi plasmid. This recombinant RNAi plasmid was checked by using the digestion of restriction enzyme and DNA sequencing techniques. Keywords mucormycosis myosin RNAi Mucor circinelloides. 1. Introduction Mucormycosis zygomycosis is the rare but lethal fungal infection especially in immunocompromised patients such as those with diabetes organ-transplanted AIDS hematologic malignancies or trauma .
