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An efficient method for constructing a random insertional mutant library for forward genetics in
Nannochloropsis oceanica - Journal of Oceanology and Limnology Vol. 42, Issue 1, Pages: 216-225(2024)
- Affiliations：
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
- Author bio：
qdguoli@139.com
yguanpin@ouc.edu.cn
- Funds：
DOI：10.1007/s00343-023-2332-1
CLC：Received：27 September 2022，
Accepted：12 January 2023，
Online First：17 March 2023，
Published：03 January 2024
- Accepted：
Scan QR Code
ZHANG Zhongyi,LIU Hang,PAN Xiaohui,et al.Research Paper An efficient method for constructing a random insertional mutant library for forward genetics in ,Nannochloropsis,oceanica[J].Journal of Oceanology and Limnology,2024,42(01):216-225.
ZHANG Zhongyi,LIU Hang,PAN Xiaohui,et al.Research Paper An efficient method for constructing a random insertional mutant library for forward genetics in ,Nannochloropsis,oceanica[J].Journal of Oceanology and Limnology,2024,42(01):216-225. DOI： 10.1007/s00343-023-2332-1.
摘要
Abstract
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
Keywords
references
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