1.Cloning Expression and Functional Verification of recF and recO Gene from Deinococcus radiodurans R1 in Escherichia coli
Lan YANG ; Xiaosong CHANG ; Wenjuan FU
Journal of Environment and Health 2007;0(08):-
Objective To construct expressing recombinant of recF and recO from Deinococcus radiodurans R1and express the target protein in E.coli BL21(DE)3.Methods recF and recO gene were amplified by PCR from genomic DNA of Deinococcus radiodurans R1 and inserted into expression plasmid vector pET30b(+) to construct pET30b(+)-recF and pET30b(+)-recO.The recombinant plasmids were transformed into E.coli BL21(DE)3 and the recombinant proteins were expressed by the isopropyl-?-D-thiogalactopyranoside(IPTG) and were analyzed with SDS-PAGE.The changes in the survival rate of bacteria in each group before-and after UV-irradiation were calculated.Results The recombinant plasmid pET30b(+)-recO and pET30b(+)-recF was obtained and the recombinant protein could be highly expressed in E.coli BL21(DE)3.Conclusion This study has provided a foundation for further studies and applications of the recombinant RecF and RecO,and initial detection shows that recO and recF gene can increase the resistant ability of E.coli.
2.Gene sequence analysis on CO1 and ND1 of Echinococcus g ranulosus
Jingfeng YU ; Wei TAN ; Bin LI ; Xiaosong LIU ; Jianhua CHANG ; Xiaoye YANG ; Rui WANG ; Lianru YANG ; Xiuxia LI
Chinese Journal of Zoonoses 2014;(8):812-815
The purpose of this study was to clarify the strains of E .granulosus from sheep of Xiwuqi and people of Xil-inhot in Inner Mongolia region and its genotypes .CO1 and ND1 of mitochondria gene were cloned and sequenced ,and then they were analyzed by MegAlign of DNAStar5 .0 .Results showed that the length of CO1 and ND1 gene of E .granulosus ,which were from sheep of Xiwuqi or people of Xilinhot ,were 936 bp and 895 bp ,respectively .The homology of CO1 gene sequences of E .granulosus strains from Xiwuqi and Xinjiang was 99 .3% ,while the homology of the corresponding gene sequences of E .granulosus from man of Xilinhot City and Xinjiang were 98 .6% .ND1 gene of E .granulosus of sheep from Xiwuqi and hu-man from Xilinhot were identical to ND1 gene of G1 type .All these indicated that the homology of E .granulosus from the two regions were high and the genotype were G1 type ,which provided an important basis for the determination of strains ,and it al-so had a great significance to prevent and control the disease .
3.Spatiotemporal Dynamics of the Molecular Expression Pattern and Intercellular Interactions in the Glial Scar Response to Spinal Cord Injury.
Leilei GONG ; Yun GU ; Xiaoxiao HAN ; Chengcheng LUAN ; Chang LIU ; Xinghui WANG ; Yufeng SUN ; Mengru ZHENG ; Mengya FANG ; Shuhai YANG ; Lai XU ; Hualin SUN ; Bin YU ; Xiaosong GU ; Songlin ZHOU
Neuroscience Bulletin 2023;39(2):213-244
Nerve regeneration in adult mammalian spinal cord is poor because of the lack of intrinsic regeneration of neurons and extrinsic factors - the glial scar is triggered by injury and inhibits or promotes regeneration. Recent technological advances in spatial transcriptomics (ST) provide a unique opportunity to decipher most genes systematically throughout scar formation, which remains poorly understood. Here, we first constructed the tissue-wide gene expression patterns of mouse spinal cords over the course of scar formation using ST after spinal cord injury from 32 samples. Locally, we profiled gene expression gradients from the leading edge to the core of the scar areas to further understand the scar microenvironment, such as neurotransmitter disorders, activation of the pro-inflammatory response, neurotoxic saturated lipids, angiogenesis, obstructed axon extension, and extracellular structure re-organization. In addition, we described 21 cell transcriptional states during scar formation and delineated the origins, functional diversity, and possible trajectories of subpopulations of fibroblasts, glia, and immune cells. Specifically, we found some regulators in special cell types, such as Thbs1 and Col1a2 in macrophages, CD36 and Postn in fibroblasts, Plxnb2 and Nxpe3 in microglia, Clu in astrocytes, and CD74 in oligodendrocytes. Furthermore, salvianolic acid B, a blood-brain barrier permeation and CD36 inhibitor, was administered after surgery and found to remedy fibrosis. Subsequently, we described the extent of the scar boundary and profiled the bidirectional ligand-receptor interactions at the neighboring cluster boundary, contributing to maintain scar architecture during gliosis and fibrosis, and found that GPR37L1_PSAP, and GPR37_PSAP were the most significant gene-pairs among microglia, fibroblasts, and astrocytes. Last, we quantified the fraction of scar-resident cells and proposed four possible phases of scar formation: macrophage infiltration, proliferation and differentiation of scar-resident cells, scar emergence, and scar stationary. Together, these profiles delineated the spatial heterogeneity of the scar, confirmed the previous concepts about scar architecture, provided some new clues for scar formation, and served as a valuable resource for the treatment of central nervous system injury.
Mice
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Animals
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Gliosis/pathology*
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Cicatrix/pathology*
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Spinal Cord Injuries
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Astrocytes/metabolism*
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Spinal Cord/pathology*
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Fibrosis
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Mammals
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Receptors, G-Protein-Coupled