产谷氨酰胺转胺酶菌株的筛选及其产酶条件研究

利用设计的低廉简便的凝胶法从土壤中分离得到1株高产谷氨酰胺转胺酶(microbialtransglutami-nase,MTG)的菌株,初步鉴定属于放线菌纲的链霉菌属(Streptomycessp.)。摇瓶发酵试验表明,其最适产酶氮源和碳源分别是蛋白胨和葡萄糖,通过正交试验确定最适产酶培养基为(g/L):葡萄糖20.0,蛋白胨20.0,酵母提取物3.0,MgSO4·7H2O2.0,K2HPO4·3H2O2.0,KH2PO42.0,CaCO35.0。单因素试验确定最佳培养时间和最适pH分别为72h和7.0;在优化条件下,发酵液中MTG酶活达1.04U/mL。     

Effect of dipeptide on intestinal peptide transporter 1 gene expression: An evaluation using primary cultured chicken intestinal epithelial cells

Peptide transporter 1 (PepT1) is a transporter responsible for absorbing dipeptide and tripeptide in enterocytes and is upregulated by dipeptide in mammals. It has not been certain whether intestinal PepT1 expression is responsive to dipeptides in chickens because of the lack of in vitro study using the cultured enterocytes. This study established a primary culture model of chicken intestinal epithelial cells (IECs) in two-dimensional monolayer culture using collagen gel by which the response of chicken PepT1 gene expression to dipeptide stimuli was evaluated. The cultured chicken IECs showed the epithelial-like morphology attached in a patch-manner and exhibited positive expression of cytokeratin and epithelial cadherin, specific marker proteins of epithelial cells. Moreover, the chicken IECs exhibited the gene expression of intestinal cell type-specific marker, villin1, mucin 2, and chromogranin A, suggesting that the cultured IECs were composed of enterocytes as well as goblet and enteroendocrine cells. PepT1 gene expression was significantly upregulated by synthetic dipeptide, glycyl-l-glutamine, in the cultured IECs. From the results, we herein suggested that dipeptide is a factor upregulating PepT1 gene expression in chicken IECs.     

前处理对大豆分离蛋白热凝胶流变学特性的影响

为了研究不同因素对大豆分离蛋白热凝胶流变学特性所产生的影响,通过改变大豆分离蛋白液的加热温度、pH以及无机盐的含量,采用旋转流变仪研究了不同大豆分离蛋白热凝胶的流变学特性.结果表明,大豆分离蛋白热凝胶的弹性模量G'随着扫描频率的降低而减小;制备凝胶时的热处理温度、盐离子含量以及pH均对大豆分离蛋白热凝胶的流变学特性产生...     

乌苏里拟鲿(Pseudobagrus ussuriensis)同工酶分析

采用聚丙烯酰胺凝胶电泳方法,对黑龙江水系乌苏里拟鲿(Pseudobagrus ussuriensis)心、肝、肾、眼、肌肉和性腺共6种组织进行了9种同工酶(ADH、EST、G6PDH、GDH、IDH、LDH、MDH、POD、SOD)的分析。结果表明,乌苏里拟鲿EST、GDH、IDH、POD同工酶谱中存在不同程度的组织特异性,EST表现出明显的性别差异性。共记录19个基因位点,多态位点百分数P=21.05%,种群遗传偏离指数d=-1,该乌苏里拟鲿种群内部杂合子缺失较严重,偏离Hardy-Weinberg平衡,遗传多样性较低。     

DNA recovery from agarose gels with a simple centrifuge-driven sephadex filtration

Conventional methods of DNA recovery from agarose gel generally require expensive equipment, extended elution times, or considerable handling of the sample after elution. We developed a simple protocol for a quick and effective recovery of DNA from agarose gels with good yield and quality. Using a Sephadex resin filled spin column, DNA fragments of 500 bp to 6 kb in an agarose gel slice were easily recovered by a 2 min centrifugation. The recovery efficiencies were over 40%-50% and the eluted DNA can be used directly for downstream application, such as polymerase chain reactions (PCR) and restriction enzyme digestion. This method could also be used to recover large DNA fragment (48 kb) without degradation. The use of Sephadex helps to remove small molecular impurities from agarose and it also reduces the chance of clogging the column filter caused by direct contact with agarose.     

干旱胁迫对白桦实生幼苗叶片蛋白质的影响

The studies on leave protein of Betula platyphylla seedlings by the methods of SDS-PAGE electrophoresis and dimensional gel electrophoresis was concerned. The result indicated that some new proteins such as 29kD pI4.3, 29kD pI4.2, 14kD pI5.3 and 11.5kD pI5.16 protein point were induced by soil drought stress, and some protein disappeared,for instance four protein between 12.4 - 11.5kD pI5.8 - 6.44 in seedlings leaves of B. platyphylla during soil drought stress.