An efficient method for constructing a random insertional mutant library for forward genetics in Nannochloropsis oceanica  

作　　者：Zhongyi ZHANG Hang LIU Xiaohui PAN Yanan ZONG Leili FENG Lixian LIU Li GUO Guanpin YANG 

机构地区：[1]College of Marine Life Sciences,Ocean University of China,Qingdao 266003,China [2]Institutes of Evolution and Marine Biodiversity,Ocean University of China,Qingdao 266003,China [3]Key Laboratory of Marine Genetics and Breeding of Ministry of Education,Ocean University of China,Qingdao 266003,China

出　　处：《Journal of Oceanology and Limnology》2024年第1期216-225,共10页海洋湖沼学报（英文）

基　　金：the National Key R&D Program of China(Nos.2018YFD0901506,2018YFD0900305);the Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology(Qingdao)(No.2018 SDKJ0406-3)。

摘　　要：Insertional mutation,phenotypic evaluation,and mutated gene cloning are widely used to clone genes from scratch.Exogenous genes can be integrated into the genome during non-homologous end joining(NHEJ)of the double-strand breaks of DNA,causing insertional mutation.The random insertional mutant library constructed using this method has become a method of forward genetics for gene cloning.However,the establishment of a random insertional mutant library requires a high transformation efficiency of exogenous genes.Many microalgal species show a low transformation efficiency,making constructing random insertional mutant libraries difficult.In this study,we established a highly efficient transformation method for constructing a random insertional mutant library of Nannochloropsis oceanica,and tentatively tried to isolate its genes to prove the feasibility of the method.A gene that may control the growth rate and cell size was identified.This method will facilitate the genetic studies of N.oceanica,which should also be a reference for other microalgal species.

关 键 词：Nannochloropsis oceanica genetic transformation random insertional mutant library zeocin pretreatment forward genetics 

分 类 号：Q78[生物学—分子生物学]