人神经母细胞瘤的磷酸化膜蛋白质组分析

针对人神经母细胞瘤SH-SY5Y细胞系的磷酸化膜蛋白质组,发展了基于多酶酶解法结合杂化硅胶基质固定化钛离子亲和色谱(Ti 4+-IMAC)整体柱富集的分析策略。该方法通过对细胞裂解液进行超速离心,以及1 mol/LNaCl和0.1 mol/L Na2CO3顺序清洗,获得膜蛋白质组分。所提取的蛋白质分别经胰蛋白酶、胰凝乳蛋白酶和胃蛋白酶平行酶解,产生的肽段经Ti 4+-IMAC整体柱选择性富集磷酸肽后,采用纳升级反相液相色谱分离和质谱鉴定,成功鉴定到43个磷酸化蛋白质,其中有14个定位于膜上。研究结果表明,采用该策略开展SH-SY5Y细胞系磷酸化膜蛋白质组学分析有望加速对该肿瘤的研究和相关潜在标记物的筛选。

Prediction Model of Secondary Substances in Anthocyanins Synthesis of Purple Corn

The aim of this study was to assay the polyphenols,flavonoid,polyphenol oxidase and phenylalnine ammonialyase which were relative to the anthocyanins synthesis of purple corn. The optimization of multiple linear regression model of anthocyanins synthesis was y=4.383 86-0.205 45x1＋5.479 638x2＋0.195 575x4. According to standard partial regression coefficient testing,the result indicated that polyphenols content was negatively correlated with anthocyanins and the relative influence to anthocyanins synthesis was-42.7%; flavonoid content and activity of polyphenol oxidase were positively correlated with anthocyanins of purple corn and the relative influence to anthocyanins synthesis were 71.45% and 73.32% respectively. There was no positive correlation between the activity of phenylalnine ammonialyase and anthocyanins of purple corn. The establishment of multiple linear regression model of anthocyanins synthesis was to provide theory foundation of producing anthocyanins in laboratory.

基于响应面法对香菇原生质体制备条件的优化

香菇Lentinula edodes是我国食用菌年产量最大的单品,优化香菇原生质体的制备条件,获得高质量、高活力的原生质体可以为香菇育种和遗传多样性分析提供技术保障。本研究利用单因素试验对稳渗剂浓度、酶解液类型、酶解时间、酶解温度4个因素的最优作用条件和相关性进行检验和筛选;依据单因素试验的结果,对酶解时间、酶解温度、酶解液种类进行响应面法3因素3水平试验优化分析。单因素试验结果表明,使用2%的溶壁酶+蜗牛酶+纤维素酶混合酶制剂为酶解液、0.6mol/L甘露醇为稳渗剂的效果显著高于其他处理(P<0.05)。经Box-Behnken设计得出香菇原生质体最佳制备条件为酶解时间3.25h,酶解液浓度1.7%,酶解温度30.49℃,获得原生质体产量为14.02×10^(6)CFU/mL,与理论产量(13.862×10^(6)CFU/mL)偏差小,可为后续原生质体融合育种和多样性分析提供重要基础。