松口蘑菌丝体的分离和RAPD-PCR分析

针对松口蘑 [Tricholomamatsutake(S .ItoetImai)Sing .]菌丝体分离培养困难和各种相关分离物目前难以用出菇试验鉴定的现实 ,采用 8种培养基配方 ,对 9个不同来源的松口蘑子实体的不同部位及菌根、菌土进行组织分离 ,计接种试管 81 0多支 ,结果从菌褶部位获得 94支慢生型的菌丝体分离菌株 ,从菌柄部位仅获得 1支快生型的菌丝体分离菌株。以马铃薯葡萄糖土壤滤液培养基 (PDAS)、马铃薯葡萄糖麦麸滤液培养基 (PDAW )、BM培养基、马铃薯葡萄糖琼脂培养基 (PDA)对菌褶进行组织分离 ,获慢生型菌丝体的成功率依次为 74.4%、35.5%、156%和 8.9%。以各分离菌株的来源松口蘑子实体和中日两国松口蘑研究者提供的分离菌株作为DNA参照样品 ,对从供试子实体、菌根、菌土进行组织分离获得的各种相关纯培养物进行亲菌鉴定。采用筛选的 1 7个随机引物介导 2 5个供试松口蘑子实体及其分离菌体的RAPD(RandomAmplifiedPolymorphicDNA) PCR反应 ,全部获得了清晰而稳定的DNA指纹图谱 ,结果一致表明 :每个松口蘑子实体的菌盖 (含菌褶…     

颈髓损伤后线粒体系列酶活性变化与线粒体功能的关系

为了探讨颈髓损伤后颈髓线粒体系列酶活性变化与线粒体功能的关系,采用Ａｌｅｎ法造成猫颈髓损伤,观察颈髓损伤后线粒体Ｃａ２＋,Ｍｇ２＋－ＡＴＰ酶、Ｎａ＋,Ｋ＋－ＡＴＰ酶、超氧化物歧化酶（ＳＯＤ）活性及线粒体呼吸功能的变化。结果显示：颈髓损伤后２ｈ至７２ｈ,Ｃａ２＋,Ｍｇ２＋－ＡＴＰ酶、Ｎａ＋,Ｋ＋－ＡＴＰ酶活性、ＳＯＤ活性明显降低,而线粒体呼吸控制率（ＲＣＲ）、磷氧比值（Ｐ／Ｏ）、氧化磷酸化效率（ＯＰＲ）也明显下降。表明颈髓损伤后Ｃａ２＋,Ｍｇ２＋－ＡＴＰ酶、Ｎａ＋,Ｋ＋－ＡＴＰ酶、ＳＯＤ活性与线粒体功能密切相关,提示颈髓线粒体的病理生理改变在颈髓损伤后继发性损害过程中起重要作用。     

EFFECTS OF INCREASED TEMPERATURE AND CO2 ON PHOTOSYNTHESIS,GROWTH, AND ELEMENTAL RATIOS IN MARINE SYNECHOCOCCUS AND PROCHLOROCOCCUS (CYANOBACTERIA)1

Little is known about the combined impacts of future CO₂ and temperature increases on the growth and physiology of marine picocyanobacteria. We incubated Synechococcus and Prochlorococcus under present‐day (380 ppm) or predicted year‐2100 CO₂ levels (750 ppm), and under normal versus elevated temperatures (+4°C) in semicontinuous cultures. Increased temperature stimulated the cell division rates of Synechococcus but not Prochlorococcus. Doubled CO₂ combined with elevated temperature increased maximum chl a–normalized photosynthetic rates of Synechococcus four times relative to controls. Temperature also altered other photosynthetic parameters (α, Φ_max, E_k, and ) in Synechococcus, but these changes were not observed for Prochlorococcus. Both increased CO₂ and temperature raised the phycobilin and chl a content of Synechococcus, while only elevated temperature increased divinyl chl a in Prochlorococcus. Cellular carbon (C) and nitrogen (N) quotas, but not phosphorus (P) quotas, increased with elevated CO₂ in Synechococcus, leading to ～20% higher C:P and N:P ratios. In contrast, Prochlorococcus elemental composition remained unaffected by CO₂, but cell volume and elemental quotas doubled with increasing temperature while maintaining constant stoichiometry. Synechococcus showed a much greater response to CO₂ and temperature increases for most parameters measured, compared with Prochlorococcus. Our results suggest that global change could influence the dominance of Synechococcus and Prochlorococcus ecotypes, with likely effects on oligotrophic food‐web structure. However, individual picocyanobacteria strains may respond quite differently to future CO₂ and temperature increases, and caution is needed when generalizing their responses to global change in the ocean.     

Analysis of variable sites between two complete South China tiger (Panthera tigris amoyensis) mitochondrial genomes

In order to investigate the mitochondrial genome of Panthera tigris amoyensis, two South China tigers (P25 and P27) were analyzed following 15 cymt-specific primer sets. The entire mtDNA sequence was found to be 16,957 bp and 17,001 bp long for P25 and P27 respectively, and this difference in length between P25 and P27 occurred in the number of tandem repeats in the RS-3 segment of the control region. The structural characteristics of complete P. t. amoyensis mitochondrial genomes were also highly similar to those of P. uncia. Additionally, the rate of point mutation was only 0.3% and a total of 59 variable sites between P25 and P27 were found. Out of the 59 variable sites, 6 were located in 6 different tRNA genes, 6 in the 2 rRNA genes, 7 in non-coding regions (one located between tRNA-Asn and tRNA-Tyr and six in the D-loop), and 40 in 10 protein-coding genes. COI held the largest amount of variable sites (9 sites) and Cytb contained the highest variable rate (0.7%) in the complete sequences. Moreover, out of the 40 variable sites located in 10 protein-coding genes, 12 sites were nonsynonymous.     

Effects of macrophage-specific adiponectin expression on lipid metabolism in vivo

Epidemiological studies have associated low circulating levels of the adipokine adiponectin with multiple metabolic disorders, including metabolic syndrome, obesity, insulin resistance, type II diabetes, and cardiovascular disease. Recently, we reported that adiponectin selectively overexpressed in mouse macrophages can improve insulin sensitivity and protect against inflammation and atherosclerosis. To further investigate the role of adiponectin and macrophages on lipid and lipometabolism in vivo, we engineered the expression of adiponectin in mouse macrophages (Ad-TG mice) and examined effects on plasma lipoproteins and on the expression levels of genes involved in lipoprotein metabolism in tissues. Compared with the wild-type (WT) mice, Ad-TG mice exhibited significantly lower levels of plasma total cholesterol (-21%, P < 0.05) due to significantly decreased LDL (-34%, P < 0.05) and VLDL (-32%, P < 0.05) cholesterol concentrations together with a significant increase in HDL cholesterol (+41%, P < 0.05). Further studies investigating potential mechanisms responsible for the change in lipoprotein cholesterol profile revealed that adiponectin-producing macrophages altered expression of key genes in liver tissue, including apoA1, apoB, apoE, the LDL receptor, (P < 0.05), and ATP-binding cassette G1 (P < 0.01). In addition, Ad-TG mice also exhibited higher total and high-molecular-weight adipnection levels in plasma and increased expression of the anti-inflammatory cytokine IL-10 as well as a decrease in the proinflammatory cytokine IL-6 in adipose tissue. These results indicate that macrophages engineered to produce adiponectin can influence in vivo gene expression in adipose tissue in a manner that reduces inflammation and macrophage infiltration and in liver tissue in a manner that alters the circulating lipoprotein profile, resulting in a decrease in VLDL and LDL and an increase in HDL cholesterol. The data support further study addressing the use of genetically manipulated macrophages as a novel therapeutic approach for treatment of cardiometabolic disease.     

花生种子高纯度DNA的提取（简报）

花生种子高纯度ＤＮＡ的提取（简报）王艳,何军贤,傅家瑞（中山大学生命科学学院,广州５０２７５）关键词花生;种子;ＤＮＡ;十六烷基三甲基溴化铵ＲＡＰＩＤＡＮＤＥＦＦＩＣＩＥＮＴＰＵＲＩＦＩＣＡＴＩＯＮＯＦＤＮＡＦＲＯＭＰＥＡＮＵＴ（ＡＲＡＣＨＩＳＨＹＰ...     

人骨保护素(OPG)重组腺病毒的制备及其生物活性研究

采用RT-PCR法得到人OPG的编码区cDNA,克隆至穿梭质粒pShuttle,构建重组有OPG编码区cDNA的腺病毒DNA,经Pac I 酶切线性化,在脂质体介导下转染HEK293细胞,制备重组腺病毒并测定病毒滴度约为5×10⁶～1.5×10⁷ pfu/mL。体外感染小鼠成肌细胞C2C12,Western blot及ELISA检测证实有OPG蛋白的表达,并可在细胞培养上清中持续表达6周。感染OPG重组腺病毒的C2C12细胞生长状态良好、细胞周期无明显变化。将重组腺病毒加入体外培养的小鼠骨髓细胞的培养基中,诱导形成的破骨细胞数量及在象牙片上形成的吸收陷窝的数量显著减少（P<0.01）。      

热球菌目(Thermococcales)遗传操作系统研究与应用进展

热球菌目(Thermococcales)是一类分离自浅海热泉或者深海热液口的超嗜热微生物,包括火球菌(Pyrococcus)、热球菌(Thermococcus)、古老球菌(Palaeococcus)。研究其生命活动的分子机制,基因的功能等必须借助遗传操作系统。由于选择标记的限制,Thermococcales遗传操作系统落后于其他菌株。近年来,在Thermococcales发现了内源质粒并可以将其改造用作遗传工具。如在Thermococcus kodakarensis及Pyrococcus furious等菌株内都建立了成熟的遗传系统,并用于基因敲除以及基因表达。将就Thermococcales内源质粒的发现和遗传操作系统的发展与应用加以阐述。