基因编辑婴儿的伦理、法律和社会蕴含

 同试管婴儿、克隆人、商业化代孕、干细胞治疗等医疗新技术引发的伦理争议相比，贺建奎基因编辑婴儿事件引发的关注更大、社会反响更为猛烈。探讨贺建奎基因编辑婴儿事件引发国际国内热议的原因，讨论了基因编辑婴儿的伦理、法律和社会蕴含。建议明确界定基因编辑技术的适用对象和范围，建立健全新型医疗技术临床应用的伦理审查和监管体系，提升全社会的伦理意识。     

植物基因环境效应启动子

植物的生长发育受到光照、温度、水分及氧等环境因素的影响，根据对不同环境的响应，植物基因的启动子主要可以分为光效应启动子，温度效应启动子和水效应启动子。在这些启动子中，除了含有基本启动子外，还存在一些特异的顺式调节元件，这些顺式调节元件在特异表达中发挥重要作用。     

大鼠肝ADAMs种类鉴定及肝再生过程中ADAMsmRNA差异分析

以 2 / 3肝切除 (patialhepatectomy ,PH)大鼠为材料 ,利用PT -PCR技术 ,通过ADAMs通用引物初步分析了肝脏中ADAMs种类及PH后恢复 4h和 36h时ADAMsmRNA差异。结果表明 :PH后不同恢复时间ADAMsmRNA的扩增谱带没有差异 ,但扩增谱带的强弱有变化 ;cDNA克隆和测序分析表明 ,大鼠肝脏有ADAM15和ADAM 19mRNA同源序列。     

噬菌体展示禽流感病毒抗原变异性基因片段文库的构建

构建T7噬菌体展示禽流感病毒抗原变异性基因片段文库. 首先， 从Gene Bank中查找筛选禽流感病毒抗原变异性基因, 将其截短、 修饰、 简并后得到禽流感病毒抗原变异性基因微阵列. 其次， 将合成的禽流感病毒抗原变异性基因片段文库扩增、 酶切, 链接到双酶切后的T7噬菌体载体基因上, 构成重组噬菌体DNA. 最后， 重组噬菌体DNA经体外包装和扩增, 得到T7噬菌体展示文库, 并进行T7噬菌体展示文库滴度、 重组率和免疫活性测定. 实验结果表明, 从Gene Bank中查找、 筛选、 剪切和修饰共获得96 258条序列构建T7噬菌体展示文库, 原始文库滴度为3.6×107个菌落/mL, 重组率大于90%. 用禽流感病毒H5N1抗体进行捕获, 经聚合酶链式反应(PCR)鉴定, 得到理想目的条带, 证明噬菌体表面展示蛋白具有抗原活性, 可用于禽流感病毒感染患者的快速检测及抗原表位筛选.     

Specific anti-tumor effect induced by attenuated Salmonella typhimurium vaccine expressing extracellular region of vascular endothelial growth factor receptor 2

The purposes of this research were to study the stable expression of exogenous gene encoding therapeutic protein in attenuated Salmonella typhimurium, observe the metabolism of oral gene vaccine carried by attenuated Salmonella typhimurium in BALB/c mouse, and investigate the feasibility of prevention and treatment of tumors by the recombinant bacteria. Recombinant plasmid pcDNA3.1 VEGFR2(n1-7) was transformed into competent attenuated Salmonella typhimurium SL3261 to develop oral DNA vaccine SL3261-pcDNA3.1 VEGFR2(n1-7). To observe whether the exogenous gene can be expressed in the recombinant bacteria, PCR was performed to amplify the CMV promoter of the eukaryotic expression vector as the proof of stable expression of exogenous protein; transmission elec- tron microscopy (TEM) was applied to observe the morphology of the recombinant bacteria to confirm that the exogenous gene has no impact on the growth of the bacteria, and then BALB/c mice were immunized with the gene vaccine. After inoculation of the gene vaccine, the recombinant bacteria SL3261 could be detected in the tissues such as small intestine, colon, liver and spleen. And then, mice in each group were challenged with tumor cells. The results of animal experiment showed that tumor growth of the mice in experimental group was inhibited and survival time of immunized mice was prolonged compared with control groups. A higher lymphocyte infiltration in tumors from animals treated with DNA vaccine was observed. Immunohistochemical analysis of tumor samples revealed an en- hanced accumulation of CD8 cytotoxic T lymphocytes, as well as an increase in CD4 cells in the tumors of animals treated with the oral gene vaccine compared to tumors from control group mice. Ultrastructure of the tumor tissue showed that tumor cells in the samples of the immunized mice were well-differentiated. Our research confirmed that the exogenous gene can be stably expressed in the attenuated Salmonella typhimurium and has no impact on the growth of the recombinant bacteria; the exogenous gene can de delivered to the host by attenuated Salmonella typhimurium to produce anti-tumor effect with no obvious cytotoxity to the host. In this study, it is established that attenuated Salmonella typhimurium could be used as a vector for oral gene vaccine, and our study provided a theoretical basis for the body distribution and the metabolism of the recombinant bacteria. This strategy may provide a simple, safe and effective way for the prevention and treatment of tumors.     

Finding finer functions for partially characterized proteins by protein-protein interaction networks

Based on high-throughput data, numerous algorithms have been designed to find functions of novel proteins. However, the effectiveness of such algorithms is currently limited by some fundamental factors, including (1) the low a-priori probability of novel proteins participating in a detailed function; (2) the huge false data present in high-throughput datasets; (3) the incomplete data coverage of functional classes; (4) the abundant but heterogeneous negative samples for training the algorithms; and (5) the lack of detailed functional knowledge for training algorithms. Here, for partially characterized proteins, we suggest an approach to finding their finer functions based on protein interaction sub-networks or gene expression patterns, defined in function-specific subspaces. The proposed approach can lessen the above-mentioned problems by properly defining the prediction range and functionally filtering the noisy data, and thus can efficiently find proteins’ novel functions. For thousands of yeast and human proteins partially characterized, it is able to reliably find their finer functions (e.g., the translational functions) with more than 90% precision. The predicted finer functions are highly valuable both for guiding the follow-up wet-lab validation and for providing the necessary data for training algorithms to learn other proteins.     

钙调神经磷酸酶B亚基高表达条件及产业化研究

重组人CNB高表达大肠杆菌已由本室构建成功,摇瓶发酵液表达量可达100mg·L-1. 通过改进培养基配方及摇瓶表达工艺,使CNB在摇瓶发酵液中表达量达到500mg*L-1以上.在高表达量的基础上,又进行了产业化改进,以蒸馏水添加无机盐取代了原配方中的自来水,并提高了表达量.     

基因工程菌株EscherichiacoliC600（pRLK14）的流加培养

在１Ｌ搅拌发酵罐中对含有重组质粒ｐＲＬＫ１４的大肠杆菌进行了培养。结果表明，采用二段培养法，于３４℃增殖细菌，在对数增殖后期升温到４２℃进行诱导，可以高效表达基因产品半乳糖激酶。诱导期，醋酸浓度增长较快，模拟实验表明，控制醋酸浓度可进一步提高半乳糖缴酶产率。     

Molecular cloning and expression of interleukin Ibeta (IL-1β) from red seabream (Pagrus major)

The interleukin 1β (IL-1β) cDNA was cloned from the red seabream (Pagrus major) by homology cloning strategy. A cDNA fragment was amplified by PCR using two degenerated primers, which were designed according to the conserved regions of other known IL-1β sequences, and elongated by 3' ends and 5' ends RACE PCR to get the full length coding sequence of red seabream IL-1β (RS IL-1β). The sequence contained 1252 nucleotides that included a 5' untranslated region (UTR) of 84bp, a 3' UTR of 410 bp and an open reading frame (ORF) of 759 nucleotides which could be translated into a putative peptide of 253 amino acids with molecular weight of 28.6 kD and putative isoelectric point pI of 5.29. The deduced peptide contained two potential N-glycosylation sites and an identifiable IL1 family signature, but lacked the signal peptide and the clear ICE cut site, which were common in other nonmammalian IL-1β genes. The RS IL-1β had the highest homology with piscine IL-1β according to phylogenetic tree analysis.     

白血病小鼠模型中P53、C-myc基因表达的变化研究

探讨DNA、RNA病毒诱导小鼠发生白血病的机制。用单纯疱疹病毒Ⅱ型（HSV－2）和C型RNA病毒诱导70只昆明小鼠，光学显微镜检测HSV－2、C型病毒诱导发生白血病的成功率。正常对照组为30只昆明小鼠。免疫组化法检测HSV－2、C型病毒致瘤后P^53、G－myc基因表达的变化。70只昆明小鼠中有47只发生白血病，成功率为67．1％。47只白血病小鼠P^53、C－myc蛋白阳性表达率均比正常对照组高（P＜0．05）；23只未形成白血病的实验组小鼠P^53、C－myc蛋白阳笥表达率均与正常对照组无显差异（P＞0．05）。HSV－2、C型病毒可以通过调节P^53、C－myc基因的表达从而诱导小鼠白血病的发生。