Objective： To clone and construct the recombinant plasmid containing ATP synthase lipid-binding protein-like protein gene of Schistosomajaponicum,（SjAslp） and transfer it into mammalian cells to express the objecti...Objective： To clone and construct the recombinant plasmid containing ATP synthase lipid-binding protein-like protein gene of Schistosomajaponicum,（SjAslp） and transfer it into mammalian cells to express the objective protein. Methods： By polymerase chain reaction （PCR） technique, SjAslp was amplified from the constructed recombinant plasmid pBCSK＋/SjAslp, and inserted into cloning vector pUCm-T. Then, SjAslp was subcloned into an eukaryotic expression vector pcDNA3.1（＋）. After identifying it by PCR, restrictive enzymes digestion and DNA sequencing, the recombinant plasmid was transfected into HeLa cells using electroporation, and the expression of the recombinant protein was analyzed by immunocytochemical assay. Results： The specific gene fragment of 558 bp was successfully amplified. The DNA vaccine of SjAslp was successfully constructed. Immunocytochemical assay showed that SjAslp was expressed in the cytoplasm of HeLa cells. Conclusion： SjAslp gene can be expressed in eukaryotic system, which lays the foundation for development of the SjAslp DNA vaccine against schitosomiasis.展开更多
Distinct phospholipid species display specific distribution patterns across cellular membranes,which are important for their struc-tural and signaling roles and for preserving the integrity and functionality of the pl...Distinct phospholipid species display specific distribution patterns across cellular membranes,which are important for their struc-tural and signaling roles and for preserving the integrity and functionality of the plasma membrane and organelles.Recent advance-ments in lipid biosensor technology and imaging modalities now allow for direct observation of phospholipid distribution,trafficking,and dynamics in living cells.These innovations have markedly advanced our understanding of phospholipid function and regulation at both cellular and subcellular levels.Herein,we summarize the latest developments in phospholipid biosensor design and applica-tion,emphasizing the contribution of cutting-edge imaging techniques to elucidating phospholipid dynamics and distribution with unparalleled spatiotemporal precision.展开更多
基金the Scientific Research Foundation of Education Committee of Hunan (07C713 and 07C708)
文摘Objective： To clone and construct the recombinant plasmid containing ATP synthase lipid-binding protein-like protein gene of Schistosomajaponicum,（SjAslp） and transfer it into mammalian cells to express the objective protein. Methods： By polymerase chain reaction （PCR） technique, SjAslp was amplified from the constructed recombinant plasmid pBCSK＋/SjAslp, and inserted into cloning vector pUCm-T. Then, SjAslp was subcloned into an eukaryotic expression vector pcDNA3.1（＋）. After identifying it by PCR, restrictive enzymes digestion and DNA sequencing, the recombinant plasmid was transfected into HeLa cells using electroporation, and the expression of the recombinant protein was analyzed by immunocytochemical assay. Results： The specific gene fragment of 558 bp was successfully amplified. The DNA vaccine of SjAslp was successfully constructed. Immunocytochemical assay showed that SjAslp was expressed in the cytoplasm of HeLa cells. Conclusion： SjAslp gene can be expressed in eukaryotic system, which lays the foundation for development of the SjAslp DNA vaccine against schitosomiasis.
基金supported by the National Natural Science Foundation of China(92354305,32321004,and 91957106 to K.H.)the Ministry of Science and Technology of the People’s Republic of China(2022YFA1304500 to K.H.).
文摘Distinct phospholipid species display specific distribution patterns across cellular membranes,which are important for their struc-tural and signaling roles and for preserving the integrity and functionality of the plasma membrane and organelles.Recent advance-ments in lipid biosensor technology and imaging modalities now allow for direct observation of phospholipid distribution,trafficking,and dynamics in living cells.These innovations have markedly advanced our understanding of phospholipid function and regulation at both cellular and subcellular levels.Herein,we summarize the latest developments in phospholipid biosensor design and applica-tion,emphasizing the contribution of cutting-edge imaging techniques to elucidating phospholipid dynamics and distribution with unparalleled spatiotemporal precision.
