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四脊滑螯虾造血组织细胞培养条件的优化
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作者 刘林敏 阮灵伟 +1 位作者 黄智豪 施泓 《应用海洋学学报》 CAS CSCD 北大核心 2024年第2期236-244,共9页
稳定的体外细胞培养体系,可以为基因功能和宿主-病原相互作用研究提供平台。本研究通过对四脊滑螯虾(Cherax quadricarinatus)造血组织(hematopoietic tissue, HPT)细胞的体外培养条件进行优化,以期获得状态稳定、活力良好的造血组织细... 稳定的体外细胞培养体系,可以为基因功能和宿主-病原相互作用研究提供平台。本研究通过对四脊滑螯虾(Cherax quadricarinatus)造血组织(hematopoietic tissue, HPT)细胞的体外培养条件进行优化,以期获得状态稳定、活力良好的造血组织细胞培养物。我们从四脊滑螯虾中分离提取造血组织细胞后,通过对培养基种类(L-15、Express Five、DMEM),谷氨酰胺浓度(0、1%、5%、10%),胎牛血清浓度(0、1%、5%、10%),以及体外培养温度(20、27℃)等培养条件进行优化。细胞形态观察和细胞活力分析结果表明,HPT细胞在添加1%青霉素-链霉素、10%L-谷氨酰胺、1%胎牛血清的L-15培养基中状态最佳,在20℃培养温度下能存活30 d左右。这些培养条件将有利于四脊滑螯虾造血组织细胞的体外培养。 展开更多
关键词 海洋生物学 造血组织细胞 原代细胞 优化培养 四脊滑螯虾
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深海热液区管状蠕虫Ridgeia piscesae新型C型凝集素的克隆与表达
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作者 陈燕婕 阮灵伟 +1 位作者 施泓 陈新华 《应用海洋学学报》 CSCD 北大核心 2023年第2期195-202,共8页
C型凝集素作为凝集素家族的主要成员之一,在无脊椎动物中有着丰富的多样性,其独特的生物学功能广泛应用于生物、医药等诸多领域。本研究从采集于东北太平洋深海热液区胡安·德富卡洋脊的管状蠕虫Ridgeia piscesae中克隆获得了一种新... C型凝集素作为凝集素家族的主要成员之一,在无脊椎动物中有着丰富的多样性,其独特的生物学功能广泛应用于生物、医药等诸多领域。本研究从采集于东北太平洋深海热液区胡安·德富卡洋脊的管状蠕虫Ridgeia piscesae中克隆获得了一种新型C型凝集素基因RPCL。该基因与已报道的凝集素的相似性都很低,并且三维结构预测有异于传统凝集素,提示其可能具有一些特殊的功能性质。进一步的原核表达成功实现了RPCL的异源重组表达,通过透析复性获得了大量可溶性蛋白,为其相关功能性质的研究打下了坚实的基础。 展开更多
关键词 海洋生物学 凝集素 管状蠕虫Ridgeia piscesae 克隆 表达
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Molecular characteristics of three thymosin-repeat proteins from Marsupenaeus japonicus and their responses to WSSV infection 被引量:5
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作者 MA Jinyou ruan lingwei +1 位作者 XU Xun GAO Zhaoming 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2016年第4期44-50,共7页
β-thymosins, a family of highly conserved peptides, play a vital role in wound-healing, angiogenesis,antimicrobial process and antiviral immunity. Three novel β-thymosin-repeat proteins, named mjthm4, mjthm3 and mjt... β-thymosins, a family of highly conserved peptides, play a vital role in wound-healing, angiogenesis,antimicrobial process and antiviral immunity. Three novel β-thymosin-repeat proteins, named mjthm4, mjthm3 and mjthm2, were cloned from Marsupenaeus japonicus using expressed sequence tags(EST) from suppression subtractive hybridization. The mjthm4, mjthm3 and mjthm2 c DNAs possessed open reading frames that encoded166, 128 and 90 amino acid residue polypeptides and contained four, three and two β-thymosin actin binding modules, respectively. Blast analysis demonstrated that mjthm4, mjthm3 and mjthm2 shared high homology with known invertebrate multi-repeat β-thymosins. These proteins are ubiquitously expressed in all of the examined tissues, and the transcriptional levels were highest in the intestine. Further investigation revealed that mjthm4,mjthm3 and mjthm2 were remarkably up-regulated 6 h after WSSV infection. Moreover, while mjthm4 transcriptional levels displayed no changes, mjthm3 and mjthm2 levels decreased in the virus-resistant shrimps.The results indicate that mjthm4, mjthm3 and mjthm2 are novel multi-repeat β-thymosin homologues, have a close relationship with WSSV infection, and might contribute to a better understanding of host defense and/or virus invasion interactions in shrimps. 展开更多
关键词 β-thymosin thymosin-repeat proteins white spot syndrome virus(WSSV) Marsupenaeus japonicus
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Characterization of four hemocyanin isoforms in Litopenaeus vannamei 被引量:4
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作者 XU Jingxiang ruan lingwei +4 位作者 LI Zhen YU Xiaoman LI Sedong SHI Hong XU Xun 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2015年第2期36-44,共9页
In this study, the gene encoding hemocyanin subunit L, Lv Hc L, was cloned from Litopenaeus vannamei and the genomic organization was characterized. This gene was diverse with many SNPs and also had at least four isof... In this study, the gene encoding hemocyanin subunit L, Lv Hc L, was cloned from Litopenaeus vannamei and the genomic organization was characterized. This gene was diverse with many SNPs and also had at least four isoforms, while one of them(Lv Hc L4) only had two exons and the exon2 was missed. Transcription analysis showed that these isoforms of Lv Hc L were up-regulated after WSSV challenge in WSSV-resistant shrimp, while the transcriptions were decreased constantly in WSSV-susceptible shrimp. It is suggested that the hemocyanin had rich polymorphism and was involved in the antiviral response. These results could extend our previous findings and provide insights into the immune feature of hemocyanin, which would be helpful for further studies aimed at antiviral mechanism in invertebrate. 展开更多
关键词 hemocyanin SNPs isoforms Litopenaeus vannamei WSSV
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A novel beta-galactose-specific lectin of the tubeworm,Ridgeia piscesae,from the hydrothermal vent
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作者 ruan lingwei XU Haipeng +3 位作者 LIN Wenyang SHI Hong CUI Zhizhong XU Xun 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2017年第6期61-67,共7页
Lectins are sugar-specific binding proteins or glycoproteins that play important physiological roles in cellular recognition and regulation. And they are also valuable in medicine and pharmacy. Tubeworm is the represe... Lectins are sugar-specific binding proteins or glycoproteins that play important physiological roles in cellular recognition and regulation. And they are also valuable in medicine and pharmacy. Tubeworm is the representative species around the hydrothermal vent in the deep sea. They have developed unique mechanisms to adapt to the harsh environment. In this study, a 1 092 bp cDNA, designed as rpgal, was first cloned and characterized from the tubeworm Ridgeia piscesae. Sequence analysis showed that RPGAL had low homology with the known galectin. And it had two homologous carbohydrate-recognition domains, which is the characteristic of the tandem-repeat type galectins. The RPGAL was successfully recombinant expressed in Escherichia coli and purified. Analysis of biological activity revealed that RPGAL was metal ion independent and it could agglutinate all the vertebrate erythrocytes tested. It was stable at 10-50℃ and pH 5-10. And the hemagglutinating activity of RPGAL was strongly inhibited by D-Lactose and lipopolysaccharide. Although RPGAL had no effect on the microorganisms tested, it showed anti-tumor activity towards HeLa cells and HT1080 cells, which was accomplished by apoptosis. The study demonstrated that RPGAL was a novel galectin and provided a potential candidate for therapy of anti-tumor. 展开更多
关键词 GALECTIN ANTITUMOR APOPTOSIS tubeworm Ridgeia piscesae
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