Expression of the Phycoerythrin Gene of Gracilaria lemaneiformis (Rhodophyta) in E. coli and Evaluation of the Bioactivity of Recombinant PE

Phycoerythrin (PE) is one of the most important proteins involved in light capturing during photosynthesis in red algae. Its potential biological activities had gained wide concerns. In the present study, tumor cytotoxic and hydroxyl radical assay were preformed to detect the bioactivity of recombinant PE. Recombinant plasmids pGEX-PE and pBGL were transformed into E.coli BL21 to make two recombinant strains BEX (pGEX-PE) and BGL (pBGL). PE expressing in BEX (pGEX-PE) was validated by SDS-PAGE and Western blotting analysis. SDS-PAGE analysis indicated that the PE-GST fusion protein was mostly inclusion bod- ies. Specific expression of PE was confirmed by Western blotting analysis. The recombinant E. coli BEX (pGEX-PE) cells were col- lected and sonicated. The supernatants were reserved for the tumor cytotoxic experiments. The result of tumor cytotoxic assay indi- cated that the supernatants containing PE had the activity of inhibiting the growth of Hela cells and with the increase of protein con- centration, the inhibiting rate increased from 37.31% to 63.26%, which showed significant difference from the control. Hydroxyl radical scavenging effect was tested with supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates treated with sonication and heating. For the sonication samples, the scavenging rates of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were significantly higher than the negative control BL21(pGEX-4T) (P<0.02), and the scavenging rates increased slowly following the increase of the protein content. For the heating samples, except for the 0.2 mg mL-1 BGL (pBGL) products, the scavenging effects of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were stronger than that of negative control BL21(pGEX-4T). However, the effect intensity was not positively correlated with the increase of the protein concentration. Though a partially de- creased hydroxyl radical scavenging activity was led by heating, the biological activity was still retained and conspicuous. This re- search showed that phycoerythrin protein expressing in E. coli has the potential medical and sanitarian value.     

莆田近岸海域海带烂孔病起因的初步研究

2019年3月莆田近岸海域海带尾部烂孔后逐渐向根部蔓延,孔洞相连后海带脱落,时空规律为先南后北逐渐发病,共涉及养殖面积约7 km2,给莆田近岸海带养殖带来了较大的损失。初步研究分析了海带大规模烂孔的原因,主要结果如下:海区温度异常升高,盐度持续较低,连续阴天光照不足,环境变化超出了细胞生理承受能力从而造成海带烂孔。此次诊断,对渔业生产及海洋环境监测结果在渔业上的应用均具有一定的现实意义。     

6-磷酸山梨醇脱氢酶(gutD)基因克隆、测序和表达

本研究根据6-磷酸山梨醇脱氢酶基因(gutD)两端序列设计引物，以假单胞杆菌(Pseudonmonas XM)的总DNA为模板通过PCR的方法克隆gutD基因并进行序列分析，将克隆得到的gutD基因与原核表达载体pBV220连接并转化大肠杆菌(Escherichia coli)JM101后检测其蛋白表达及菌株生长耐盐性。     

一株深海耐嗜压菌的细胞毒活性次生代谢产物研究

通过CCK8法检测分离自深海一株深海耐压菌Shewanella piezotolerans WP3次生代谢产物的细胞毒活性,研究了温度、压力、噬菌体对WP3次生代谢产物细胞毒活性的影响.发现WP3次生代谢产物具有良好细胞毒活性.低温因素可提高其细胞毒活性,在100μg/cm^3的含量下,代谢产物对肿瘤细胞Bel7402、CNE2、A549、HT1080细胞毒活性均有提高,其中对A549细胞毒活性提高了1.3倍.在最适温度（20℃）情况下,高压可降低其细胞毒活性.20 MPa培养条件下,代谢产物对4株肿瘤细胞均未表现出明显活性,而常压培养条件下对SW 480具有93.3%的抑制率,对HT1080抑制率表现出87.5%,对CNE2抑制率是79.8%.在低温情况下,压力因素对其细胞毒活性无影响,噬菌体对细胞毒活性影响不大.HPLC指纹图谱分析发现不同温度条件下,其次生代谢产物指纹图谱有变化,结合细胞毒活性结果分析,为后期分离鉴定细胞毒活性化合物提供指导.     

福建闽南地区与广西北海地区波纹巴非蛤的形态差异与遗传多样性分析

本研究采用形态学和微卫星分子标记对福建闽南地区（厦门、云霄、惠安）及广西北海4个野生波纹巴非蛤群体进行形态学与遗传多样性分析。主成分与聚类分析结果表明：惠安与北海群体的形态最为相近,其次为云霄群体,而厦门群体的趋异度最大。构建了两个主成分PC1和PC2,其中PC1的贡献率为36.134%,PC2的贡献率为21.443%,累计贡献率为57.577%。建立了4群体的判别函数,判别准确率P₁为43.3%~93.5%,P₂为44.8%~88.0%,综合判别率为66.1%。4个波纹巴非蛤群体的整体遗传多样性较高,群体间遗传分化微弱,属于同1个自由交配组群,NJ系统发育树聚类结果未检测到与地理位置相对应的谱系结构,这可能与人为引种、贸易及增殖放流等活动导致波纹巴非蛤不同群体间频繁基因交流有关。建议今后在波纹巴非蛤的资源管理和开发利用时,加强本地种质资源的保护。     

广西珍珠湾红树种群结构与动态特征

本研究以种群生命表为基础,对位于广西珍珠湾内的桐花树(Aegiceras corniculatum)种群、白骨壤(Avicennia marina)种群、秋茄(Kandelia obovata)种群和木榄(Bruguiera gymnorrhiza)种群采用径级法及匀滑技术,编制种群特定时间生命表,绘制种群存活曲线、死亡率和消失率曲线,进行种群数量动态变化和时间序列分析。结果表明:(1)白骨壤种群、秋茄种群和木榄种群年龄结构呈倒"J"形,种群趋于Deevey-II型,即增长型,主要集中在I龄级,存活曲线、死亡率和消失率曲线均呈现出更新层死亡率较高的特征;(2)桐花树种群年龄结构呈"金字塔"形,属于Deevey-I型,为衰退型,更新层个体数量少,种群总体不具备显著增长性幅度;(3)4种红树种群的数量变化动态指数V_pi和V′_pi均大于0.00%,种群稳定但易受外部环境影响;(4)在未来2、4、6、8个龄级时间后,4种红树种群中老龄级个体能够得到不同程度的补充。     

大亚湾西部海域中国毛虾种群动态变化及其对环境要素的响应

本研究依据2016年12月至2017年2月和2017年12月至2018年3月的每月上、中、下旬在大亚湾西部海域进行的5条拖网测线的中国毛虾(Acetes chinensis)调查,以及2016年10—12月浮游植物光合色素的调查,分析了中国毛虾的种群动态变化及其对主要环境因子的响应.结果表明:中国毛虾总个体密度在L2和...     

广西涠洲岛滨海湿地潮下带海草、红树林与互花米草的分布和群落结构特征

系统报道了广西涠洲岛的潮下带海草、红树林以及互花米草的分布和群落结构特征.2019—2020年的调查结果表明,涠洲岛海草分布于南湾西侧,紧邻珊瑚礁区,面积4246 m2,为潮下带的生长形式,海草种类仅见卵叶喜盐草(Halophila ovalis);真红树植物有6种[红海榄(Rhizophora stylosa)、海漆...     

鸟蛤苗种培育及增养殖技术

通过对鸟蛤（Cardium sp．）的生物学研究,进行了苗种培育、稚贝中间培育、养成、增殖技术的综合系统研究,探明了鸟蛤的生态分布、食性、繁殖习性、生长发育、埋栖及移动生态等生物学特性,报道了鸟蛤人工苗种培育及增养殖技术措施,系统地提出了鸟蛤苗种控温培育及增养殖技术工艺流程与操作规范。