DNA vaccination against foot-and-mouth disease via electroporation: study of molecular approaches for enhancing VP1 antigenicity

BACKGROUND: Foot-and-mouth disease virus (FMDV) affects susceptible livestock animals and causes disastrous economic impact. Immunization with plasmid expressing VP1 that contains the major antigenic epitope(s) of FMDV as cytoplasmic protein (cVP1) failed to elicit full protection against FMDV challenge. MATERIALS AND METHODS: In this study, mice were immunized via electroporation with four cDNA expression vectors that were constructed to express VP1 of FMDV, as cytoplasmic (cVP1), secreted (sVP1), membrane-anchored (mVP1) or capsid precursor protein (P1), respectively, to evaluate whether expression of VP1 in specific subcellular compartment(s) would result in better immune responses. RESULTS: Electroporation enhanced immune responses to vectors expressing cVP1 or P1 and expedited the immune responses to vectors expressing sVP1 or mVP1. Immunization of mice via electroporation with mVP1 cDNA was better than sVP1 or cVP1 cDNA in eliciting neutralizing antibodies and viral clearance protection. Vaccination with P1 cDNA, nonetheless, yielded the best immune responses and protection among all four cDNAs that we tested. CONCLUSIONS: These results suggest that the antigenicity of a VP1 DNA vaccine can be significantly enhanced by altering the cellular localization of the VP1 antigen. Electroporation is a useful tool for enhancing the immune responses of vectors expressing VP1 or P1. By mimicking FMDV more closely than that of transgenic VP1 and eliciting immune responses favorably toward Th2, transgenic P1 may induce more neutralizing antibodies and better protection against FMDV challenge.     

Ap、Km和Cm抗性细菌在土壤中的差异分布及成因

从不同区域土壤中分离细菌：对其进行氨苄青碡素、卡那霉素和氯霉素的抗性测试,以及抗药性质粒分析。结果表明,对照区和生活区土壤细菌抗氨苄青霉素和卡那霉素菌株比例未见显著性差异,未检出抗氯霉素细菌。保护区中抗氨苄青霉素、卡那霉素菌株含质粒比例均为18.2％,生活区中抗氨苄青霉素和卡那霉素菌株含质粒比例分别为36.4％和27．3％。随机抽提质粒转化无抗忡菌,表明部分抗菌素抗性基因是由质粒携带。实验结果认为本地土壤中抗菌素抗性菌分布差异尚未有显著性表现。     

细胞工程教学的实践与探索

细胞工程是高等院校生物技术专业的主干课程之一,授课教师必须明确课程的主要任务和要求,准确把握教学内容,选择灵活多样的教学方法,兼顾课堂教学和实验教学,激发学生学习兴趣,为其全面系统地掌握本专业的相关知识奠定基础。     

Screening of binding proteins that interact with human Salvador 1 in a human fetal liver cDNA library by the yeast two-hybrid system

Salvador promotes both cell cycle exit and apoptosis through the modulation of both cyclin E and Drosophila inhibitor of apoptosis protein in Drosophila. However, the cellular function of human Salvador (hSav1) is rarely reported. To screen for novel binding proteins that interact with hSav1, the cDNA of hSav1 was cloned into a bait protein plasmid, and positive clones were screened from a human fetal liver cDNA library by the yeast two-hybrid system. hSav1 mRNA was expressed in yeast and there was no self-activation and toxicity in the yeast strain AH109. Twenty proteins were found to interact with hSav1, including HS1 (haematopoietic cell specific protein1)-associated protein X-1 (HAX-1); neural precursor cell expressed, developmentally down-regulated 9, pyruvate kinase, liver and RBC, cytochrome c oxidase subunit Vb, enoyl coenzyme A hydratase short chain 1, and NADH dehydrogenase (ubiquinone) 1 beta subcomplex, demonstrating that the yeast two-hybrid system is an efficient method for investigating protein interactions. Among the identified proteins, there were many mitochondrial proteins, indicating that hSav1 may play a role in mitochondrial function. We also confirmed the interaction of HAX-1 and hSav1 in mammalian cells. This investigation provides functional clues for further exploration of potential apoptosis-related proteins in disease biotherapy.     

基于CSSL的高密度物理图谱定位水稻分蘖角度QTL

对以籼稻9311为遗传背景携带粳稻日本晴基因组的染色体片段置换系（CSSL）的遗传图谱进行分子标记加密,构建了含250个多态标记的高密度物理图谱。以119个CSSLs为材料,P≤0.001为阈值,筛选到分蘖角度与受体亲本9311差异极显著的10个系。结合物理图谱和代换作图方法,共鉴定出5个分蘖角度QTL,其中qTA11的加性效应表现为增效作用,来源于9311的等位基因;其余4个QTL的加性效应为减效作用,均来源于日本晴的等位基因。qTA6-1和qTA6-2分别被定位于第6染色体RM253–RM527之间的3.55Mb区段和RM3139–RM494的1.65Mb区间;qTA9被定位于第9染色体RM257–RM189之间的3.40Mb区段;qTA10被定位在第10染色体RM222–S10-1之间的2.10Mb区段;qTA11被定位于第11染色体RM1761–RM4504之间的3.30Mb区间。以上研究结果为水稻分蘖角度QTL的精细定位和株型育种提供了依据。     

伤寒沙门菌与甲、乙、丙副伤寒沙门菌质控血清的制备

应用免疫学原理,将伤寒沙门菌O901、H901和甲、乙、丙型副伤寒沙门菌分别制成全菌体抗原,免疫实验兔获取免疫血清。依据伤寒沙门菌和副伤寒沙门菌的抗原成分的异同性,选择适当的吸收菌除去免疫血清中的交叉反应抗体和类属凝集素,而保留其特异性的抗体。通过对诊断菌液的验证试验,证实吸收充分的免疫血清具有质控血清的特性。具备可靠性能的质控血清,适用于伤寒沙门菌与副伤寒沙门菌的菌种检定及其效价检测;亦有利于肥达氏诊断菌液的质量控制。     

甘露糖正向筛选体系在巨尾桉遗传转化中的应用

为了研究甘露糖正向筛选体系在巨尾桉遗传转化过程中的有效性,构建了以6-磷酸甘露糖异构酶（6-phosphomannose isomerase,PMI）为筛选标记的pCAMBIA1301植物表达载体,并将该载体通过农杆菌介导的遗传转化转入木本植物巨尾桉中。将获得的阳性植株通过氯酚红（chlorophenol red,CPR）法及PCR检测,桉树遗传转化的阳性率达到26.09%。另外,通过正交试验优化法,对巨尾桉组培快繁体系建立过程中不同浓度激素配比进行了研究,建立起良好的巨尾桉组织培养再生体系,由甘露糖筛选敏感性测试,获得了巨尾桉筛选临界浓度,蔗糖与甘露糖比例为19∶11,优化了巨尾桉遗传转化体系,为今后巨尾桉组织培养与遗传转化研究提供了重要的参考依据。