DNA末端硫修饰依赖的体外大片段DNA连接法

【背景】DNA组装技术是基因组合成中的一个关键技术。探索低成本、高效率的基因组合成技术一直是合成生物学的重要研究领域。在某些细菌如变铅青链霉菌中,DNA上有磷硫酰化修饰(简称硫修饰),而在另一些细菌如天蓝色链霉菌中存在一种含有硫修饰识别结构域(sulfur-binding domain, SBD)的识别蛋白,可以特异性识别DNA上的硫修饰,这启发了我们发展出一种新的DNA组装技术。【目的】探究在DNA末端硫修饰的连接中,T4 DNA连接酶与SBD相融合蛋白和单独的T4 DNA连接酶相比,是否有更高的连接效率。【方法】根据同源重组原理,设计硫修饰引物,扩增硫修饰的DNA片段。构建T4 DNA连接酶与SBD融合蛋白的3种表达载体T4-linker-SBD(Hga)、T4-linker-SBD(Spr)和T4-linker-SBD(Mmo),表达纯化以上3种融合蛋白。比较3组浓度梯度(2.4、0.24、0.024 mg/mL) T4 DNA连接酶与融合蛋白在2.5 kb和8.0 kb DNA片段连接上的差异。【结果】DNA末端硫修饰的2.5kb和8.0kb的两端片段均能扩增,而且3种融合蛋白...     

Marine biological injuries and their medical management: A narrative review

The marine environment can be extremely dangerous, and the harm caused by marine organisms when they contact the human body can be especially harmful, even deadly. Contact includes stings, bites, wounds, and consumption as food. In this article, the characteristics of the common marine biological injuries are summarized, the major marine organisms causing damage in China’s marine waters are described, and injury prevention and treatment methods are discussed.     

SEQUENCES OF LLANDOVERIAN CHITINOZOANS IN YANGZl REGION

This paper is dedicated as a token of homage and gratitude to Prof. Mu. En-zhi, the nestor in the field of graptolite studies, who introduced us chitinozoan studies with constant supports in many ways, and who died on 8th April, 1987. The Palaeontological Society of China has decided to publish a specieal volume in memory of this outstanding scientist.     

人结核分枝杆菌特异性核酸探针制备及结核病PCR检测技术的初步临床应用

采用人结核分枝杆菌(Mycobacterium tuberculosis TB)染色体DNA为模板,选择位于插入片段IS6110中884～865和568～588碱基对处的两个片段为引物,扩增出317bp的特异性片段．将其克隆进pUCl9载体。酶切图谱分析和DNA序列测定证实为目的片段。该片段经DIG标记,分别与11种分枝杆菌DNA进行Southern杂交,结果证明只与人型复合分枝杆菌发生杂交反应。利用该对引物建立的PcR检测拄术对74份结核病痰液标本进行检测,并与临床细菌快速培养结果相比较,发现48份临床阳性均为PcR阳性,在26份临床阴性标本中亦发现11份PCR检测阳性。将标本PCR产物与克隆探针进行杂交,显示两者结果完全一致。说明PCR检测体系结果可靠,其灵敏度明显高于目前临床所采用的方法,可作为一种常规技术用于结核病的临床检测。     

背角无齿蚌晶杆的扫描电镜观察

本文应用扫描电镜的方法,对背角无齿蚌的晶杆进行了研究。结果表明：晶杆外形似圆柱状,可分为二部分,即前端膨大的头部和其后渐细的杆状部。前者分左右两瓣,略弯曲,与杆状部形成一个锐角,表面呈“蜂窝状”,似一种膜性结构,其上吸附有大量的消化酶和粘液;后者两侧有凹槽,表面具许多凹陷小坑,多为圆形或卵圆形,大小不一,深浅不同,且排列不规则。未凹陷处也粗糙不平。晶杆的功能主要是搅拌和消化食物。同时,本文对晶杆结构与功能的关系也进行了讨论。     

异色瓢虫若虫对麦三叉蚜的捕食作用

研究表明,异色瓢虫高龄若虫捕食麦二叉蚜的功能反应为ＨｏｌｌｉｎｇⅡ型反应,喜好捕食低龄若蚜;在４０℃下捕食率最低,１０℃次之,２５℃最高．在１０～２５℃内捕食率ｙ与温度ｘ的关系为ｙ＝２８．１３０３＋２．６６６５ｘ,在２５～４０℃内为ｙ＝１９９．９２７５－３．６４２ｘ;在种内干扰条件下,其捕食作用率Ｅ与开敌密度Ｐ的关系为Ｅ＝０．６８９７Ｐ－０．７９３０,搜索常数Ｑ为０．６８９７,干扰系数ｍ为０．７９３０;其分摊竞争强度Ｉ与异色瓢虫密度Ｐ的关系为Ｉ＝０．０４００＋１．０３８３ｌｏｇＰ．     

猪骨胶原蛋白酶解物中血管紧张素转换酶抑制剂的提纯

将猪骨胶原蛋白粗提物用胰蛋白酶水解,经阳离子交换树脂层析,ＳｅｐｈａｄｅｘＧ－２５柱凝胶过滤,以及数次反相高效液相层析,最终获得一具有抑制血管紧张素转换酶（ＡＣＥ）活性的单一峰值的多肽。其氨基酸组成为Ｉｌｅ,Ｈｉｓ,Ｓｅｒ,Ｇｌｙ,Ａｌａ,Ｐｒｏ,Ｔｙｒ,Ｌｅｕ,Ａｓｐ．以Ｈｉｐ－Ｇｌｙ－Ｇｌｙ为底物,在ｐＨ为７．１的条件下,此肽对猪肺ＡＣＥ的Ｉ＿（５０）值为２６μｍｏｌ／Ｌ。     

中国大陆若干群体的黑果蝇的线粒体DNA多态性研究

本文研究了果蝇Ｄ．ｖｉｒｉｌｉｓ种群Ｄ．ｖｉｒｉｌｉｓ线粒体ＤＮＡ（ｍｉｔｏｃｈｏｎｄｒｉａｌＤＮＡ,ｍｔＤＮＡ）的多态性。用９种限制性内切酶ＸｂａⅠ,ＥｃｏＲⅠ,ＰｓｔⅠ,ＨｉｎｄⅢ,ＢｇｌⅡ,ＳａｃⅠ,ＳｃａⅠ,ＥｃｏＲＶ和ＰｕｖⅡ,对青岛、南京、上海、宁波与泉州５个Ｄ．ｖｉｒｉｌｉｓ群体的ｍｔＤＮＡ进行了限制性片段长度多态性（ｒｅｓｔｒｉｃｔｉｏｎｆｒａｇｍｅｎｔｓｌｅｎｇｔｈｐｏｌｙｍｏｒｐｈｉｓｍ,ＲＦＬＰ）的研究。在５群体中,发现５种不同的酶切图谱,它们彼此之间的遗传差异π为０．４６％－１．７６％,群体内遗传差异πｉｊ为０．００％－０．３３％,群体间的差异ｄｘｙ,为０．００％－０．８２％。分布于中国大陆的Ｄ．ｖｉｒｉｌｉｓ的群体间遗传差异在总遗传差异中所占比例γｓｔ值为２４．６２％。我们发现,Ｄ．ｖｉｒｉｌｉｓ的栖息环境对ｍｔＤＮＡ的遗传变异有十分明显的影响,而不同地理纬度的群体之间其遗传距离并无倾群（ｃｌｉｎｅ）表现。     

Protective Effects of Pyridoxal Phosphate Against Glucose Deprivation-Induced Damage in Cultured Hippocampal Neurons

Abstract: When hippocampal cultures were deprived of glucose, massive release of lactate dehydrogenase (LDH), an indicator of neuronal death, occurred via NMDA receptor activation. Addition of pyridoxal phosphate (PLP; 1 and 10 µ M ) inhibited this LDH release in a concentration-dependent manner. Prior exposure to PLP evoked more potent inhibitory effects on LDH release compared with those treated at the onset of glucose deprivation. Furthermore, PLP inhibited the reduction of intracellular content of pyruvate induced by glucose deprivation, which was accompanied by the reversal of intracellular ATP depletion. A noteworthy elevation of extracellular glutamate in response to glucose deprivation was completely reversed by addition of PLP. Aminooxyacetic acid, a potent inhibitor of PLP-dependent enzymes, antagonized the effects of PLP on LDH release, pyruvate production, and ATP formation. These results suggest that PLP protects neurons from glucose deprivation-induced damage by enhancing the formation of energy-yielding products and relieving extracellular load of glutamate. The observed phenomena further indicate that PLP might be used prophylactically against neuronal death induced by metabolic disorders.