小蜡果皮水溶性色素的提取及光敏感性

以去离子水和不同体积分数乙醇从小蜡果皮中浸提红色素,用光谱扫描法检测该红色素的光吸收特性,比较不同浸提时间及不同体积分数乙醇对浸提效果的影响,并用暴露方式进行光敏感性测定。结果显示,小蜡果皮色素在纯水中的溶解性最好,属水溶性色素,延长浸提时间可提高色素的浸出率,但杂质质量分数也随之提高,紫外线对色素的色度影响最大,直接照射可使色素的色度大幅度下降。     

Recovery and purification of plant-made recombinant proteins

Plants are becoming commercially acceptable for recombinant protein production for human therapeutics, vaccine antigens, industrial enzymes, and nutraceuticals. Recently, significant advances in expression, protein glycosylation, and gene-to-product development time have been achieved. Safety and regulatory concerns for open-field production systems have also been addressed by using contained systems to grow transgenic plants. However, using contained systems eliminates several advantages of open-field production, such as inexpensive upstream production and scale-up costs. Upstream technological achievements have not been matched by downstream processing advancements. In the past 10 years, the most research progress was achieved in the areas of extraction and pretreatment. Extraction conditions have been optimized for numerous proteins on a case-by-case basis leading to the development of platform-dependent approaches. Pretreatment advances were made after realizing that plant extracts and homogenates have unique compositions that require distinct conditioning prior to purification. However, scientists have relied on purification methods developed for other protein production hosts with modest investments in developing novel plant purification tools. Recently, non-chromatographic purification methods, such as aqueous two-phase partitioning and membrane filtration, have been evaluated as low-cost purification alternatives to packed-bed adsorption. This paper reviews seed, leafy, and bioreactor-based platforms, highlights strategies for the primary recovery and purification of recombinant proteins, and compares process economics between systems. Lastly, the future direction and research needs for developing economically competitive recombinant proteins with commercial potential are discussed.     

三七总RNA提取方法的对比研究

比较利用改进的异硫氰酸胍一步法、异硫氰酸胍高盐法、CTAB法和Thomas’RNA提取法等4种方法提取三七根茎2个部位总RNA的可行性。结果表明,改进的异硫氰酸胍一步法和异硫氰酸胍高盐法能有效地抑制酚类物质、多糖及皂苷等次级代谢产物对总RNA的影响,可从三七根茎中获得质量高、完整性好的总RNA。RT—PCR分析显示提取的总RNA具有反转录活性。这2种方法具有快速、简单、有效的特点。     

从野皂荚提胶后的副产物中制备分离蛋白

采用碱提酸沉法从野皂荚提胶后的副产物中制备分离蛋白,研究了制备工艺条件对分离蛋白得率的影响。结果表明:当料液比1∶10~1∶12(m/V)、提取液pH8.0~9.0、浸提温度45~50℃、浸提两次、每次浸提45 m in时,分离蛋白得率可达78.45%。研究结果可为野皂荚提胶副产物中蛋白的综合利用奠定了基础。     

黄酒中多糖的提取和分析

本文利用单因素和正交试验探究了黄酒中多糖提取工艺条件,并分析了黄酒多糖的化学组分。单因素实验结果表明,乙醇浓度在低于80%时,粗多糖的提取量随着乙醇浓度的增加而增加,高于80%时,粗多糖的量变化不大;醇沉时间到达第8 h时粗多糖的提取量基本达到稳定;在醇沉温度为10℃时粗多糖的量达到最大值。通过正交试验得到的黄酒多糖的最佳提取工艺为:乙醇浓度为80%,醇沉时间为6 h,醇沉温度为5℃。进一步分析纯化后多糖的化学组分为中性糖含量为89.6%、糖醛酸含量为0.48%、蛋白质含量为4%。     

一种适于转基因水稻PCR检测的微量DNA快速提取法

对已报道的小麦基因组DNA快速提取方法的部分步骤进行了简化,在水稻上进行了尝试。结果表明,简化法提取的水稻基因组DNA完整性好,PCR扩增效果与试剂盒提取法无明显的差异,结果稳定可靠;而且整个提取过程操作简单、花费时间少,样品用量少,仅需5-10mg,适用于大规模转基因水稻的PCR检测。     

猕猴桃模板DNA的提取及RAPD-PCR最佳反应体系的建立

以改良CTAB法从猕猴桃叶片中制备模板DNA ,优化了PCR热循环参数 ,建立了RAPD PCR扩增的最佳反应体系。实验结果表明 ,CTAB提取液中EDTA组分的浓度对模板提取影响很大 ,其最适浓度为 80mmol/L ;用异丙醇沉淀后不经乙醇洗涤纯化的DNA不会影响扩增效果。PCR热循环参数为 :94℃预变性 5min ;94℃变性 1min ,37℃退火 1min ,72℃延伸 2min ,循环 4 0次 ;最后在 72℃延伸 6min。     

Recent Findings in the Chemistry of Odorants from Four Baccharis Species and Their Impact as Chemical Markers

Baccharis is a widespread genus belonging to the Asteraceae family that includes almost 400 species exclusively from the Americas. Even when studied in detail, the taxonomic classification among species from this genus is not yet fully defined. Within the framework of our study of the volatile composition of the Baccharis genus, four species (B. trimera, B. milleflora, B. tridentata, and B. uncinella) were collected from the ‘Campos de Cima da Serra’ highlands of the Brazilian state of Rio Grande do Sul. The aerial parts were dried and extracted by the simultaneous distillation extraction (SDE) procedure. This is the first time that SDE has been applied to obtain and compare the volatile‐extract composition in the Baccharis genus. Characterization of the volatile extracts allowed the identification of 180 peaks with many coeluting components; these latter being detailed for the first time for this genus. The multivariate statistical analyses allowed separating the volatile extracts of the four populations of Baccharis into two separate groups. The first one included the B. milleflora, B. trimera, and B. uncinella volatile extracts. The three species showed a high degree of similarity in their volatile composition, which was characterized by the presence of high contents of sesquiterpene compounds, in particular of spathulenol. The second group comprised the extract of B. tridentata, which contained α‐pinene, β‐pinene, limonene, and (E)‐β‐ocimene in high amounts.     

Comparison of five commercial DNA extraction kits for the recovery of Yersinia pestis DNA from bacterial suspensions and spiked environmental samples

Aim:  To evaluate commercial DNA extraction kits for their ability to isolate DNA from Yersinia pestis suspensions and spiked environmental samples.
Methods and Results:  Five commercially available DNA extraction kits were evaluated: the ChargeSwitch gDNA Mini Bacteria Kit, the IT 1-2-3 Sample DNA Purification Kit, the MasterPure Complete DNA and RNA Purification Kit, the QIAamp DNA Blood Mini Kit and the UltraClean Microbial DNA Isolation Kit. The extraction methods were performed upon six Y. pestis strains and spiked environmental specimens, including three swab types and one powder type. Taqman real-time PCR analysis revealed that the use of the MasterPure kit resulted in DNA with the most consistently positive results and the lowest limit of detection from Y. pestis suspensions and spiked environmental samples.
Conclusion:  Comparative evaluations of the five commercial DNA extraction methods indicated that the MasterPure kit was superior for the isolation of PCR-amplifiable DNA from Y. pestis suspensions and spiked environmental samples.
Significance and Impact of the Study:  The results of this study can assist diagnostic laboratories with selecting the best extraction method for processing environmental specimens for subsequent detection of Y. pestis by real-time PCR.     

超临界CO2萃取红景天中红景天苷、苷元酪醇的研究

采用超临界CO₂ 萃取法和乙醇常温浸提法相比较, 研究从红景天中提取红景天苷、苷元酪醇的工艺条件, 结论是:采用超临界CO₂ 萃取法能萃取出红景天生药中红景天苷的1.2%, 提取率不高, 但该方法能萃取出80%的苷元酪醇, 萃取液中苷元酪醇的相对含量可达45.68%;乙醇常温浸提法能将红景天苷、苷元酪醇同时有效萃取, 且得率较高, 但是萃取液中两物质相对含量较低, 进一步分离纯化将有难度。本研究结果表明, 将超临界CO₂ 萃取法和乙醇常温浸提法有效结合, 可实现两物质的有效分离, 推进红景天有效成分的产业化进程。