菌根真菌液泡的分离及特性研究

从菌根真菌黑核菌（ＣｅｎｏｃｏｃｃｕｍｇｅｏｐｈｉｌｕｍＦｒ．）和鹅膏菌［Ａｍａｎｉｔａｍｕｓｃａｒｉａ（Ｌ．ｅｘＦｒ．）Ｐｅｒｓ．ｅｘＨｏｏｋ．］菌丝中，采用多元碱化合物诱导原生质体裂解方法，分离出细胞器──液泡。实验结果表明，在加磷的ＭＭＮＣ培养基中培养４—７天的菌丝经水解酶作用所释放出的原生质体具有较高的液泡化程度；液泡产率约为原生质体的５－１０％；细胞质特征酶葡萄糖－６－磷酸－脱氢酶活性测定结果证明了液泡的纯度；而等量原生质体和液泡中酸性磷酸酶及α－甘露糖苷酶活性的比较表明这两种酶主要存在于液泡中，因此，它们可以作为菌根真菌黑核菌和鹅膏菌液泡的特征酶。     

马尾松基因库无性系花期观察分析

在福建省沙县官庄林场石景山工区进行马尾松基因库无性系开花习性、花量、花期的观察。观察结果表明：不同无性系着生球花量的差异显著，始花期也有明显的差异，但也存在一定的同步性，大部分无性系雌花开放时间比雄花早４─９天。     

牛Myostatin基因单核苷酸多态性分析

Myostatin基因即肌肉生长抑制素,是一种肌肉生长的负调控因子。应运PCR-SSCP和测序的方法对中国秦川牛、南阳牛以及国外引入品种皮埃蒙特牛双肌基因的第三外显子进行了多态性分析。结果表明:第三外显子938处G→A的单核苷酸的突变造成了南阳牛、皮埃蒙特牛第三外显子扩增片段多态性,秦川牛则不然。     

Influence of the Indoleacetic Acid-Lysine Synthetase Gene (iaaL) of Pseudomonas syringae subsp. savastanoi on Yield Attributes of Potatoes

The iaaL gene of Pseudomonas syringae subsp. savastanoi encodes an indoleacetic acid-lysine synthetase which conjugates free indoleacetic acid (IAA) with lysine. lAA-lys is biologically less active than free IAA. The iaaL coding region was expressed under the control of the cauliflower mosaic virus 35S promoter and transgenic potato plants were produced (Spena et al. 1991). 35S iaaL potato plants are characterized by increased internodal length and epinastic bending of older leaves. In three greenhouse experiments with plants grown in pots of different size and in two growth chamber experiments tuber number increased in iaaL transgenic plants compared to untransformed and vector-transformed controls of the same genotype. The increase in tuber numbers observed under controlled conditions was reflected in tuber yield which increased in the pot grown transgenics.     

表达序列标签（ESTs）及其应用

表达序列标签(ESTs)是指从(ESTcDNA文库中随机挑取克隆并对其3’或5’端进行单轮自动测序所获得的短cDNA库列，一般长度为300～500bp。要有效获取ESTs，必须对cDNA文库进行预处理，处理方法有衰减杂交法、均一化法和差异显示法。在dbEST中收录了大量各种生物的ESTs。ESTs广泛应用于鉴定基因、发现新基因、电子克隆、构建遗传学图谱、制备DNA芯片、分子标记、研究基因的差异表达及检验病原微生物等方面。     

Genetic Engineering Approaches to Pig Production*

Modern biotechnology promises a number of new applications in animal breeding and production. Although conventional pig breeding has achieved a high level of efficiency and productivity numerous problems have been encountered with animal health and the loss of meat quality. Selection based on phenotypic performance data of individual animals does not take into account the importance of specific genes and their relevance within a complex regulatory system. In most cases it is therefore difficult to trace back the genetic origins of clinically important disorders. The application of genetic engineering techniques in pig production will facilitate diagnosis, improvement of productivity, and animal health by allowing direct genetic manipulation. Attention must be focussed on the physical and genetic analysis of the procine genome. The isolation and characterisation of genes, DNA-markers, polymorphic DNA-fragments, and their chromosomal assignment will be important prerequisites and tools for the elucidation of genetic disorders. Especially the detection of heterozygous carriers of recessive disorders and their elimination from the breeding stock will increase selection accuracy and decrease the generation intervals. But also the rapid and simple detection of infectious diseases, which is sometimes difficult if not impossible at present, will improve animal health and welfare. Although the production of transgenic animals either by DNA-microinjection into zygotes or the use of embryonal stem cells manipulated in vitro is less straightforward than DNA-based diagnosis it will play an important role in the direct manipulation of the porcine genome and genes. Breeding programmes including the use of transgenic livestock have already been developed. There is no doubt that genetic engineering has reached a degree of practical feasibility, allowing it to play an important role in pig breeding in particular and animal production in general.     

大白菜细胞核雄性不育基因向自交系97A407的转育

根据复等位基因遗传假说 ,利用大白菜核不育系 9810 9和已知基因型的甲型两用系可育株 99135 2作为不育源 ,以大白菜自交系 97A40 7为目标亲本 ,经过 5个世代的转育 ,获得了具有 97A40 7遗传基因型的新甲型两用系、临时保持系和雄性不育系。     

猪胸膜肺炎放线杆菌外膜蛋白(OMP)5′末端保守区基因的克隆、表达及其免疫活性研究

以猪胸膜肺炎放线杆菌血清7型25-4株基因组DNA为模板,PCR方法扩增外膜蛋白(OMP)5′末端保守区基因片段(OMPc),酶切及核苷酸序列分析鉴定后,与原核表达载体质粒pGEX-6P-1进行连接,构建成重组表达载体pGEX-OM-Pc,转入大肠杆菌BL21中,以IPTG进行诱导,SDS-PAGE电泳分析发现,转化了重组质粒的菌株所表达的融合蛋白相对分子量为34 kD,与实际预测相符,命名为GST-OMPc.GST亲和层析柱进行纯化,ELISA方法对纯化蛋白进行检测.结果表明:纯化蛋白GST-OMPc能够与兔抗猪胸膜肺炎放线杆菌血清7型的阳性血清反应.OMPc蛋白的成功表达为其功能的研究打下基础.     

新麦草YUCCA基因克隆及其表达特性分析

【目的】克隆新麦草分蘖关键基因YUCCA并明确其表达特性,为该基因功能验证和育种利用奠定基础。【方法】以不同分蘖类型的新麦草分蘖节为材料,采用PCR法克隆新麦草YUCCA基因并进行同源序列比对,构建1300-cYFP-YUCCA过表达载体,利用烟草瞬时转染及蛋白质印迹法分析新麦草YUCCA蛋白的表达位置及表达情况,通过RNA-Seq数据及qRT-PCR分析验证YUCCA基因的相对表达量。【结果】成功克隆了新麦草YUCCA基因全长,大小为1 336 bp,最大阅读框为1 236 bp,并成功构建了1300-cYFP-YUCCA过表达载体。多序列比对结果显示,新麦草YUCCA主要与麦类作物的亲缘关系较近。表达特性分析发现,YUCCA基因在多分蘖型新麦草中的相对表达量远低于少分蘖型新麦草;与叶、根等组织相比,YUCCA基因在分蘖节中的相对表达量最高。烟草瞬时转化和蛋白质印迹法分析均显示,YUCCA蛋白可以正常表达,其亚细胞定位于细胞质中。【结论】YUCCA基因在调控新麦草分蘖中起下调作用,可用于后续YUCCA基因功能的研究。