Running through the continents of Asia, Europe and Africa, the Belt and Road routes connect the vibrant East Asia economic circle, the developed European economic zone, and the encompassing countries and regions with huge potential for economic development. It will have profound and far-reaching impact on the political, economic, cultural and scientific & technological development of China in the future. As regards countries along the Belt and Road, this report analyzes the scientific and technological development foundation, studies the overall progress of biotechnology fundamental research and patent technology R&D, and summarizes the biotechnology industry development and the international patent distribution. Finally, suggestions are made accordingly to further deepen cooperation between China and its counterparts to achieve mutual benefit in the field of biotechnology.
To investigate the new genes associated with dexamethasone metabolism in order to provide an idea for the construction of an efficient and stable dexamethasone degrading engineering bacterium.
Target genes which were suspectly associated with dexamethasone metabolism were screened according to the results of the whole genome sequencing and bioinformatics analysis of the newly discovered dexamethasone degrading bacteria Burkholderia sp.CQ001 (B.CQ001). Total RNA of B.CQ001 was extracted from B.CQ001wasprocessed by RT-PCR to produce cDNA. Using cDNA as a template, the new gene was rapidly cloned subsequently by PCR technology which was verified by real-time fluorescence quantitative PCR (RT-qPCR), then connected with prokaryotic expression vector pET-28a-c (+), after DNA sequencing. The recombinant plasmid pET-28a-Ivd was transformed into B.CQ001. Using high performance liquid chromatography (HPLC) to evaluate the biodegradability of the expressed bacteria for dexmethas one sodium phosphate.
Three unknown genes were screened out from B.CQ001: ORF05499, ORF05827, ORF06535, and their expressed products were: peroxiredoxin, ketosteroid isomerase, isovaleryl coenzyme A dehydrogenase; Real time quantitative PCR analysis showed that the expression levels of three genes were upregulated in different degrees after induced by dexamethasone ,especially gene ORF06535; HPLC showed that the degradation of dexamethasone sodium phosphate and dexamethasone rate can reach to 89% and 80% after overexpression of ORF06535 in B.CQ001. Compared with the original strain B.CQ001, the degradation ability had improved significantly.
New gene ORF06535 associated with dexamethasone degradation was discovered in B.CQ001 which had been cloned and functional verified. It provides new genetic information for preparation of dexamethasone and steroid hormone bioremediation.
Genetic engineering is an important means to improve the traits of lily. To establish an efficiency and stable transformation system is the basis of transgenic research. The different regeneration systems of lily were screened. The vector containing the target gene GR and selection gene NPTII was screened by Agrobacterium tumefaciens. Establish efficiency genetic transformation system for callus and scales through orthogonal test. The results showed that the optimum culture conditions as follow. The medium of lily scale inducing was MS+2mg/L 2,4-D + 0.1mg/L NAA + 90g/L + 0.2mg/L NAA + 30g/L sucrose. Indirect differentiation medium was MS+2.5mg/L 2,4-D+0.4mg/L 0.4mg/L TDZ+60g/L sucrose. The Kana selection pressure for scales was 100mg/L and for callus was 75mg/L. The direct genetic transformation of the scales was Agrobacterium OD600 = 0.6, pre-culture for 3 days, infiltrating for 40min, As 200μmol/L. The indirect callus transformation was: Agrobacterium OD600, pre-incubated for 5 days, infiltrating for 40min, As 200μmol/L. The direct and indirect positive plant transformation rates were 17.50% and 12.60%.
To establish a qualitative and quantitative method for the determination of antibody glycosylation from the level of intact glycopeptides based on LC-MS, and to apply it to qualitative and quantitative analysis of biomimetic drugs.
The qualitative and quantitative analysis of glycoforms was carried out from the level of intact glycopeptides, and compared with the results from UPLC-FLR, the traditional glycan analysis approach. The optimal analysis strategy was established and used for the analysis of N-glycans on antibody biosimilars.
The UPLC-FLR methods obtained the quantitative result of seven glycoforms from bevacizumab; While the method of mass spectrometry quantified 19 glycoforms. After enrichment by hydrophilic interaction liquid chromatography (HILIC), 22 glycan forms were quantified. The quantitative results of the glycosylation mass of the trastuzumab showed good agreement.
Based on the method of LC-MS, it has the advantages of fast speed, high sensitivity and high detection of glycoforms, and it is expected to be a stable quantitative strategy for site - specific glycosylation analysis of biopharmaceuticals.
To develop armored RNA reference material containing target RNA of norovirus based on Qbeta bacteriophage for norovirus nucleic acid detection.
Synthesize DNA fragment named QINVGII containing maturase coding gene, capsid protein coding gene and packing site of Qbeta bacteriophage, cDNA corresponding detection target sequence of gene group II norovirus in ISO/T15216-2 2012. QINVGII fragment was cloned into prokaryotic expression vector pET-28a (+). The recombinant plasmid was identified by enzyme digestion and sequencing, and then expressed in E. coli BL21 (DE3) cells through isopropyl-β-thiogalactopyranoside (IPTG) induction. The expression product, VLPs was analyzed by SDS-PAGE and the electron microscope. The VLPs was centrifuged and purified by CsCl density gradient after digestion with DNase I and RNase A. The homogeneity and stability of the reference material were tested according to the GB/T15000.3—2008 [directives for the work of reference materials (3) - reference material -general and statistical principles for certification].
SDS-PAGE analysis showed that the molecular mass of the expressed protein product was about 14.1kDa, which was consistent with the prediction. The 25nm VLPs could be observed under electron microscope. The VLPs samples were valued as (1.06±0.06)×107 copies/μl and behaved well in the homogeneity test, F=2.24<F0.05(9,20). The stability test indicated that the sample was stable at 37℃ for 12 days, at room temperature (20~25℃) for 24 days, at 4℃ for at least 120 days, -20℃ for at least 150 days, at -80℃ for at least 300 days with no significant decrease.
The armored RNA based on Qbeta bacteriophage prepared, which had good uniformity, stability and high copy number, could be a good reference material candidate for the norovirus RNA detection.
Ebola viruses are the causative agents of Ebola hemorrhagic fever with high infection rates and high fatality rates, which were classified as category A pathogens. A safe and effective vaccine against Ebola virus infection is urgently needed. Currently, the Ebola virus vaccine candidates under study include viral vectors vaccines, protein vaccines and DNA vaccines. Among them, the most promising vaccine candidate is the recombinant vesicular stomatitis viral vector-based vaccine: rVSV-ZEBOV. The vaccine is safe and effective in prevention and treatment Ebola hemorrhagic fever. It is expected to be available in 2018. In order to gain an in-depth understanding of rVSV-ZEBOV vaccine, the preparation, pharmacological studies, and mechanism of action of rVSV-ZEBOV vaccine are focused on.
Because antimicrobial peptides have a broad spectrum of antimicrobial activity and are not easy to cause resistance, it are understood abroad multifunctional tools of the innate immune system to fight microbial infections.However, the antimicrobial activity and stability of natural antimicrobial peptides is lower, and hemolytic activity of antimicrobial peptides is higher, resulting in more difficultly applying to clinical trials. So researchers optimize to design it to achieve the newly antimicrobial peptides of higher antimicrobial activity and lower hemolytic activity. In addition, natural antimicrobial peptides are discovered as a class of immune effectors, resulting from antimicrobial activity , immunomodulatory activity and the ability of neutralizing endotoxin of antimicrobial peptides and so on. So researchers are more interested in its anti-inflammatory development. Optimized design and anti-inflammatory mechanism of antimicrobial peptides are reviewed.
Pituitary adenylate cyclase-activating polypeptide (PACAP) belongs to the secretin, glucagon, and vasoactive intestinal peptide(VIP) family. PACAP is widely distributed in the brain and peripheral organs, notably in the endocrine pancreas, gonads, respiratory and urogenital tracts. PACAP play important roles in metabolism and neuroprotection. Diabetes is a common chronic metabolic disease characterized by high blood sugar. Its complications have become the main cause of mortality and disability in diabetes patients. The knowledge accumulated over the past years on the role of this neuropeptide in diabetes and its major complications, particularly emphasizing its potential for new therapeutic approaches were outlined.
The cell viability and functions are finely regulated through various biochemical modification occurred in the metabolism process. Enzymatic glycosylation is a common molecular modification in metabolic regulation and has an important impact on maintaining and regulating cells functions. Glycosyltransferases are enzymes that catalyze the transfer glycosyl moieties from activated donor to a wide and diverse range of acceptor molecules. The glycosylation of the acceptor molecules lead to changes of their intracellular properties such as stability, solubility and regional localization, and thus played important roles in many bioprocesses including cell cycles, signal transduction, protein expression, resistance responses, and clearance of pollutants. The classification, naming and catalytic mechanism of glycosyltransferase superfamily are briefly introduced. Then the glycosylation of protein and small molecule compounds and their functions in metabolic processes are reviewed, when the glycoside was transferred to intracellular biomolecule proteins and small molecule compounds. At last,the application prospects of glycosyltransferases and glycosylation reactions in the fields of human health pharmaceutical products, industrial catalysis, food and agriculture are looked forward.
Male sterility plays an important role in hybrid vigor utilization and hybrid seed production in crops. The three-line system and two-line system based on cytoplasmic male sterility and photo-thermo sensitive male sterility had been widely used in crop hybrid seed production. But there are some limitations such as low efficiency of germplasm utilization, unstable fertility in variable environmental condition. In the last three decades, many artificial manipulations of male sterility in plants have been accomplished by using genetic engineering or biotechnology strategies. The reported approaches used for generating artificial biotechnology male-sterile lines in the main three crops such as maize, rice and wheat are outlined. Especially, detail the multi-control sterility (MCS) system in maize designed by our laboratory recently is described. This will give some insights on the commercial application of male sterility in crop breeding and hybrid seed production.
Male sterile materials play important roles both in maize heterosis utilization and hybrids production, which are based on cytobiology and molecular genetics research on maize anther development.Anther development of maize is sophisticated, which need cooperative regulation of sporophyte and gametophyte. Here, the research progresses in maize anther morphology, maize anther development division, male sterility classification, cytobiology research methods, various omics research methods and molecular genetics methods on anther development are summarized, which will provide methodological clues for understanding the mechanism underlying maize anther development and industrialized utilization of male sterile materials.
Maize (Zea mays L.) genic male-sterile mutant has been widely studied for its biological significance on pollen developmental study and commercial value in hybrid vigor utilization. As the development of molecular biotechnology, several genic male-sterile genes has been cloned and characterized in maize, leading to potential use of genetic engineering male-sterility in commercial hybrid seed production. Here, the progress of cytological characterization, gene cloning and molecular mechanism of genic male-sterility study in maize are reviewe, and potential application approaches of genic male-sterile genes in maize hybrid production is discussed.
Cytoplasmic male sterility (CMS) has become a powerful tool for hybrid production in maize,which is one of the most successful crop with heterosis utilization. CMS caused by incompatibility between cytoplasmic and nuclear gene products,and the fertility canbe restored by nuclear fertility restoration(Rf)genes. Maize has three major types of CMS, namely CMS-T, CMS-S and CMS-C, on the basis ofthe reaction to specific restorers.Advance on the CMS and Rf genes of each type is reviewed, and the application situation inmaizehybrid production is analysed.
Photo- or/and thermo-sensitive genic male sterile (P/TGMS) line can be used as a male sterile line as well as a maintainer line in different ecological environment. P/TGMS is an important resource for hybrid seed production by simplifying its procedure. The production modes of hybrid seed for major crops and reviewed the progress in P/TGMS lines and its application in hybrid seed production of rice, wheat, maize, millet, etc are introduced. Finally, the prospective application of P/TGMS is discussed.
Male sterility is a phenomenon in which the male gametophyte or reproductive organs lose their physiological functions, and the application of the male sterility significantly increased the efficiency of hybrid production. Because of the different mechanisms, male sterility can be classified into cytoplasmic male sterility, genetic male sterility unaffected by environmental conditions, photo/ thermo-sensitive sterility and chemical induced sterility, which have all been applied in the crop hybrid production by the “three lines” or “two lines” system. Here, the recent research advances of all these different pathways and its application in crop heterosis are reviewed.
