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| Establishment and Optimization of an Electroporation-Mediated Transfection System in Dwarf Surf Clam Mulinia lateralis |
| SUN Xiaohan1,2, DAI Shukun1,2, LIN Siyu1,2, GUO Xiaolin1,2, ZHAO Kaixuan1,2, YANG Zujing1,2, ZHANG Zhifeng1,2,3, QIN Zhenkui1,2 |
1. Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China;
2. Shandong Key Laboratory of Marine Seed Industry (preparatory), Ocean University of China, Qingdao 266003, China;
3. Key Laboratory of Tropical Aquatic Germplasm of Hainan Province, Sanya Oceanographic Institution, Ocean University of China, Sanya 572000, China |
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Abstract Exogenous gene introduction technology plays a pivotal role in trait improvement of economically important aquatic species, particularly shellfish. However, the limitations of related technological platforms severely restrict the elucidation of molecular mechanisms underlying critical traits and the advancement of genetic breeding. To identify suitable exogenous gene delivery methods for application in shellfish, a fluorescent reporter gene vector was constructed in the potential model shellfish species dwarf surf clam Mulinia lateralis using the ef1α gene promoter sequence, and the ptdTomato-ef1α2524 plasmid with high expression activity was screened through in vitro experiments. Subsequently, electroporation transfection was utilized to deliver the reporter plasmid into eggs of the dwarf surf clam. Electroporation parameters including voltage, pulse duration, pulse number, and plasmid mass concentration were optimized, and embryo-larval survival rates and transfection efficient were evaluated. It was found that the optimal electroporation were observed under conditions of seawater electroporation buffer, plasmid concentration 60 μg/mL, voltage 150 V, pulse duration 1 ms, and single pulse application. Under these conditions, there was transfection efficiency of (33.66±2.71)%. The results successfully realized efficient vector delivery into eggs of dwarf surf clam by electroporation, and established a shellfish-adapted electroporation technology platform. These findings provide robust technical support for molecular mechanism exploration, gene function studies, and breeding technology development in shellfish and other aquatic species, significantly promoting the advancement of gene editing technologies in mollusks.
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Received: 18 April 2025
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