扩展青霉基因组DNA五种提取方法比较

为寻找一种适合于扩展青霉基因组DNA的提取方法,比较了固体培养、液体静置培养和液体振荡培养3种培养方式的提取效果,分别采用氯化苄法、蜗牛酶法、CTAB法、SDS法及异硫氰酸胍法提取扩展青霉菌基因组DNA,经DNA质量浓度测定及电泳分析,比较5种方法的差异,并利用扩展青霉的多聚半乳糖醛酸酶(Polygalacturonase)基因内一段保守序列,设计PCR引物,扩增大小为288 bp的特异目的基因,检测其灵敏性.结果表明:固体培养的方式较适合扩展青霉基因组DNA的提取,另外5种提取方法均能提取出扩展青霉菌基因组DNA,扩增出目的条带,其中氯化苄法和蜗牛酶法所提DNA的纯度均较好,但蜗牛酶法所提DNA量较少.因此,氯化苄法是5种提取方法中最可靠的DNA提取方法,所提DNA浓度高、纯度好、结果稳定,可作为PCR反应的模板进行特异性扩增,且该方法的最低检测限大约为1.01×10-1μg/mL.     

MDH和G6PD同工酶在家鸡个体发育中的表现特征

采用不连续缓冲系统的聚丙烯酰胺凝胶电泳法(PAGE法)结合组织特异性染色技术,对家鸡不同发育阶段和不同组织器官的苹果酸脱氢酶(MDH)和葡萄糖-6-磷酸脱氢酶(G6PD)同工酶进行了测定.结果表明,MDH同工酶在家鸡体内既有阶段发育特征,又有组织器官特异性表现,且s-MDH和m-MDH的分布和表达的量上有较大差别,表现在s-MDH在谱带多少、组织分布普遍程度和酶活性高低上都强于m-MDH.认为MDH同工酶在家鸡体内主要参与该组织器官的脂肪代谢.G6PD同工酶在家鸡未成年之前各发育阶段和各组织器官只呈现酶谱痕迹,成年之后仅在肝、心、肾、睾丸、胸肌中出现1～2条谱带,显示出分布的发育阶段和组织器官局限性.     

应用绿粘帚孢霉防治杉苗立枯病

研究了应用绿粘帚孢霉(Gliocladium virens)的发酵制剂、菌粉和分生孢子及其与低量PCNB联合防治由Rhizoctcnia solani引起的杉苗立枯病,结果表明各种制剂与低量PCNB的联合防治效果都比各自单独使用的效果更好。其中发酵制剂与PCNB的联合防治效果最好。在土壤中发酵制剂增殖的水平最高,菌粉次之,分生孢子则不明显。低量PCNB在一定程度上增加了生物制剂增殖的水平。同时,R.solani的群体受到生物制剂的抑制,低于对照的群体密度。     

家鸡LDH同工酶的发生遗传学分析

采用不同连续缓冲系统的聚丙烯酰胺凝胶电泳法，结合组织特异性染色和酶谱区带活性扫描技术，对家鸡胚胎期和生后期不同生长发育阶段各组织器官的乳酸脱氢酶（ＬＤＨ）同工酶进行了测定。结果表明，鸡胚胎期Ｂ型ＬＤＨ同工酶占优势（５１．４￣９９．８％），生后期则有３种不同表现：心、眼、脑组织继续保持Ｂ亚基的优势地位；肝、肾组织呈现Ｂ型酶含量的微弱上升；胸肌则发生Ｂ型酶到Ａ型的颠倒转换。ＬＤＨ同工酶的这种表达特征是     

C_(18)键合磁珠固相萃取-液质联用检测蜂王浆中磺胺二甲嘧啶残留

[目的]建立一种应用C18键合磁珠固相萃取-液质联用检测蜂王浆中磺胺二甲嘧啶残留的方法。[方法]MCX固相萃取小柱联合C18磁珠对蜂王浆进行前处理,用HPLC-串联质谱仪对净化后样本进行检测。[结果]该方法的线性范围是5～80μg/kg,相关系数0.993 1。添加水平为5、10、20μg/kg时各种化合物的平均回收率范围为93%～104%,RSD值均不大于11.3%。[结论]该方法简单、快速,结合自动化设备,可实现蜂王浆样品中磺胺二甲嘧啶测定前处理的自动化。     

海滨木槿叶片蛋白质双向电泳体系的建立

为建立一套适合海滨木槿叶片蛋白质组学分析的双向电泳体系,以海滨木槿无性系叶片为材料,分别采用裂解液法、三氯乙酸一丙酮沉淀法、改良酚抽法分离纯化蛋白质,选用24empH3—10的IPG胶条,并对不同上样量、等电聚焦程序、平衡时间等电泳条件进行优化。结果表明：采用改良酚抽法提取蛋白质,上样量为每IPG胶条1000μg,总聚焦功率90kV·h,平衡时间15min,胶体考马斯亮蓝染色,可得到蛋白质点数目多、分辨率高的双向电泳图谱。重复性试验结果显示,该体系具有很好的稳定性及可重复性,可满足海滨木槿蛋白质组学研究的要求。     

2009—2011年云南省连旱对马铃薯生产的影响

从2009—2011年云南省国民经济和社会发展统计公报和其他资料看,虽然3年连旱对于马铃薯生产有影响,但还有一定程度的发展,但该种旱情有可能常态化,而且云南省水利设施的储水量占降雨量的比例明显偏低,因此对云南省干旱不能掉以轻心。建议加强水利建设、加大抗旱作物马铃薯等的发展,除应对可能会加重的干旱外,也充分地运用云南省的光热条件。     

Enhanced Hydrolysis of Excess Sludge by Two Surfactants

The hydrolysis efficiency of excess sludge by the two surfactants (SDS and SDBS) was investigated according to COD dissolution rate, concentrations of soluble carbohydrate and soluble protein. The results showed that the hydrolysis of excess sludge was improved by using the two surfactants. When the dosage was in low range, SCOD concentration increased significantly with dosage increasing. But the increase of SCOD was not obvious when the dosage was higher than 50 mg/g dw. SCOD concentration increased from 638.5 mg/L to 6 446.8 mg/L (SDBS) and 4 857.2 mg/L (SDS) respectively. COD dissolution rate increased from 5.8% to 37.3%(SDBS) and 30.2%(SDS) respectively. With the increase of SDS and SDBS dosage in the range of 0~150 mg/g dw, the concentrations of the soluble protein and carbohydrate increased linearly. Soluble carbohydrate increased from 3.54 mg/L to 95.56 mg/L(SDBS) and 64.20 mg/L(SDS) respectively. Soluble protein concentration increased from 11.72 mg/L to 706.30 mg/L(SDBS) and 541.08 mg/L(SDS) respectively. The concentrations of ammonia and VFA also increased with the SDS and SDBS dosage. Ammonia concentrations increased from 4.21 mg/L to 130.33 mg/L(SDBS) and 102.74 mg/L(SDS) respectively. VFA concentrations increased from 21.27 mg/L to 358.30 mg/L(SDBS) and 283.12 mg/L(SDS) respectively.     

采用CTAB与SDS法提取白术DNA的研究

[目的]获取白术中高质量基因组DNA。[方法]选取新鲜白术叶片,破碎细胞壁,用经改良的CTAB与SDS法进行DNA分离纯化提取。[结果]改良CTAB法提取的DNA OD值为1.8～1.9,较改良SDS法的OD值(1.5～1.6)高。[结论]改良CTAB法提取的DNA浓度和纯度较高,可作为模板用于后续分子标记的研究。     

Variation in seed proteins from mutagen-treated cultivars and selected lines of Vigna unguiculata L. Walp

The storage protein profiles of the seeds of two IITA cowpea cultivars (‘IT84E–124’ and ‘Vita 7’) exposed to three mutagens—sodium azide (NaN₃), ethyl methane sulphonate (EMS) and ⁶⁰Co gamma rays—and those of 18 selected M₃ lines were investigated by sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis (PAGE). The total seed protein, globulin and albumin fractions showed differences in number and intensity of subunit bands, but the least differences were found in the albumin fraction. Both high and low molecular weight bands were observed, the highest being 94.5 kDa and the lowest 12.0 kDa.‘Vita T showed less variability compared with ‘IT84E–124’, as indicated by the relative similarity indices (S.I.) of the two cultivars. The lowest S.I. of 0.200 was found among ‘IT84E–124’ lines while the lowest S.I. among ‘Vita T lines was 0.4706. A number of lines with particular traits were found to be characterized by the presence of specific polypeptide bands. This study demonstrates that induced mutation could create additional variability to supplement existing germplasm and that SDS-PAGE is a useful tool for discriminating and estimating genetic similarities among selections.