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151.
Dawei Jiang Yunchao Liu Aiping Wang Gaiping Zhang Guoyu Yang Yumei Chen Pengchao Ji Chang Liu Yapeng Song Yunfang Su Guoqiang Wang Jucai Wang Baolei Zhao Ruiguang Deng 《Biotechnology letters》2016,38(6):901-908
Objectives
To improve the expression of soluble IBDV VP2 protein by using different tagged vectors in Escherichia coli.Results
Fusion tags, Grifin, MBP, SUMO, thioredoxin, γ-crystallin, ArsC and PpiB, enhanced the expression and solubility of VP2 protein. The fusion proteins were purified by Ni–NTA chromatography, MBP-VP2 showed the highest purity about 90 %. After removing the MBP tag, VP2 self-assembled into virus-like particles, ~25 nm diam. Results from AGP suggested the recombinant IBDV VP2 protein identified by reference serum like IBDV.Conclusion
All the seven tags enhanced the expression and solubility of IBDV VP2 protein. The recombinant protein self-assembly into virus like particles and possess antigenicity as reference IBDV.152.
基因打靶方法制备乳腺生物反应器 总被引:4,自引:0,他引:4
基因打靶技术制备乳腺生物反应器,克服了显微注射法的众多缺陷,并随着第一例基因打靶家畜的诞生而成为这一研究领域的热点。本文从制备乳腺生物反应器过程中基因打靶策略、打靶细胞到打靶位点的选择等各个方面,详细分析了其中的技术难点、解决问题的对策、国内外研究进展以及应用前景。 相似文献
153.
Bin Hu Ye Guo Chunyuan Chen Qing Li Xin Niu Shangchun Guo Aijun Zhang Yang Wang Zhifeng Deng 《Cellular and molecular neurobiology》2014,34(6):905-912
The use of transplanting functional neural stem cells (NSCs) derived from induced pluripotent stem cells (iPSCs) has increased for the treatment of brain diseases. As such, it is important to understand the molecular mechanisms that promote NSCs differentiation of iPSCs for future NSC-based therapies. Sirtuin 1 (SIRT1), a NAD+-dependent protein deacetylase, has attracted significant attention over the past decade due to its prominent role in processes including organ development, longevity, and cancer. However, it remains unclear whether SIRT1 plays a role in the differentiation of mouse iPSCs toward NSCs. In this study, we produced NSCs from mouse iPSCs using serum-free medium supplemented with retinoic acid. We then assessed changes in the expression of SIRT1 and microRNA-34a, which regulates SIRT1 expression. Moreover, we used a SIRT1 inhibitor to investigate the role of SIRT1 in NSCs differentiation of iPSCs. Data revealed that the expression of SIRT1 decreased, whereas miRNAs-34a increased, during this process. In addition, the inhibition of SIRT1 enhanced the generation of NSCs and mature neurocytes. This suggests that SIRT1 negatively regulated the differentiation of mouse iPSCs into NSCs, and that this process may be regulated by miRNA-34a. 相似文献
154.
155.
Molecular Cloning, Co-expression, and Characterization of Glycerol Dehydratase and 1,3-Propanediol Dehydrogenase from Citrobacter freundii 总被引:1,自引:0,他引:1
Xianghui Qi Wenying Deng Fei Wang Qi Guo Huayou Chen Liang Wang Xiang He Ribo Huang 《Molecular biotechnology》2013,54(2):469-474
1,3-Propanediol (1,3-PD), an important material for chemical industry, is biologically synthesized by glycerol dehydratase (GDHt) and 1,3-propanediol dehydrogenase (PDOR). In present study, the dhaBCE and dhaT genes encoding glycerol dehydratase and 1,3-propanediol dehydrogenase respectively were cloned from Citrobacter freundii and co-expressed in E. coli. Sequence analysis revealed that the cloned genes were 85 and 77 % identical to corresponding gene of C. freundii DSM 30040 (GenBank No. U09771), respectively. The over-expressed recombinant enzymes were purified by nickel-chelate chromatography combined with gel filtration, and recombinant GDHt and PDOR were characterized by activity assay, kinetic analysis, pH, and temperature optimization. This research may form a basis for the future work on biological synthesis of 1,3-PD. 相似文献
156.
Lingling Xu Tongjian Li Liang Liao Huisheng Deng Xingjie Han 《Plant Systematics and Evolution》2013,299(3):603-610
Hybridization is particularly likely to occur in initial and young polyploid complexes, and interspecies hybridization between diverged species usually leads to a complicated reticulate evolution. The Ranunculus cantoniensis complex and its allied species include R. chinensis (2x), R. silerifolius var. silerifolius (2x), R. cantoniensis (4x), R. trigonus (2x), R. shuichengensis (2x), R. diffusus (4x), R. repens (4x), R. vaginatus (5x) and R. sieboldii (6x, 8x). Many morphological intermediates can be found between the members of this complex, and the relationship among members is complicated. By analyzing internal transcribed spacers and nrDNA FISH (fluorescence in situ hybridization) signals, we unraveled the phylogenetic and genetic constitution of the various taxonomic units of this complex. Haplotypes were highly separated by median-joining network analysis and at least four haplogroups emerged in which there were 11 primary haplotypes; six out of ten taxa shared haplotype 1, suggesting that haplotype 1, a variation of the primary haplotype R. chinensis, served as the pivotal genome in the complex. The pollen characteristics and electrophoretic patterns of R. vaginatus (5x) showed it to be an intermediate between R. diffusus (4x) and R. sieboldii (6x). The distribution of R. vaginatus (5x) was located at the junction of the distributions of R. diffusus (4x) and R. sieboldii (6x). Ranunculus vaginatus (5x) shared haplotypes 7 and 8 with R. diffusus (4x), and haplotypes 8 and 9 with R. sieboldii (6x). This proved that R. vaginatus (5x) emerged from hybridization between R. diffusus (4x) and R. sieboldii (6x). The results of FISH also support a hybrid origin of R. vaginatus (5x). The findings of this study clearly show that there are only eight taxa in this polyploid complex including R. chinensis (2x), R. silerifolius var. silerifolius (2x), R. trigonus (2x), R. silerifolius var. dolicanthus(2x), R. cantoniensis (4x), R. diffusus (2x), R. vaginatus (5x) and R. sieboldii (6x, 8x). These taxa associated with each other by hybridizing with the pivotal genome. Ranunculus cantoniensis (4x) and R. vaginatus (5x) arose from hybridization events between diverged species in the polyploid complex, leading to a complicated reticulate evolution. 相似文献
157.
Expression vectors of human granulocyte colony stimulating factor (G-CSG) and long acting tissue plasminogen activator (La-tPA)
in mammary gland were constructed using promoters of mouse whey acid protein gene (WAP) and sheep β-lactoglobulin gene (BLG)
with sizes of 2.6 and 5 kb respectively. Two kinds of transgenic mice of G-CSF and La-tPA were produced with microinjection.
The expression of G-CSF and La-tPA was achieved in mammary glands of transgenic mice, respectively. In order to establish
dual transgenic mice of La-tPA /G-CSF, transgenic mice carrying G-CSF and La-tPA gene characterized with specific expression
in mammary gland were mated. La-tPA/G-CSF dual transgenic mice were screened out from the hybrid offspring by Once-PCR. The
co-expression of La-tPA and G-CSF in mammary gland of the dual transgenic mice was confirmed by the milk assayed and Northern
blot analysis. Some parameters about the dual transgenic mice indicated that there were fewer litters than that of normal
mice. The ratio of dual transgenes was 46.1% in F1 generation, and offspring’s sex ratio was normal. Hence a dual transgenic
mouse model was established for the study of co-expression foreign proteins in mammary gland. 相似文献
158.
159.
Kimura H Kwan KM Zhang Z Deng JM Darnay BG Behringer RR Nakamura T de Crombrugghe B Akiyama H 《PloS one》2008,3(9):e3174
Background
Bone mass is maintained by continuous remodeling through repeated cycles of bone resorption by osteoclasts and bone formation by osteoblasts. This remodeling process is regulated by many systemic and local factors.Methodology/Principal Findings
We identified collagen triple helix repeat containing-1 (Cthrc1) as a downstream target of bone morphogenetic protein-2 (BMP2) in osteochondroprogenitor-like cells by PCR-based suppression subtractive hybridization followed by differential hybridization, and found that Cthrc1 was expressed in bone tissues in vivo. To investigate the role of Cthrc1 in bone, we generated Cthrc1-null mice and transgenic mice which overexpress Cthrc1 in osteoblasts (Cthrc1 transgenic mice). Microcomputed tomography (micro-CT) and bone histomorphometry analyses showed that Cthrc1-null mice displayed low bone mass as a result of decreased osteoblastic bone formation, whereas Cthrc1 transgenic mice displayed high bone mass by increase in osteoblastic bone formation. Osteoblast number was decreased in Cthrc1-null mice, and increased in Cthrc1 transgenic mice, respectively, while osteoclast number had no change in both mutant mice. In vitro, colony-forming unit (CFU) assays in bone marrow cells harvested from Cthrc1-null mice or Cthrc1 transgenic mice revealed that Cthrc1 stimulated differentiation and mineralization of osteoprogenitor cells. Expression levels of osteoblast specific genes, ALP, Col1a1, and Osteocalcin, in primary osteoblasts were decreased in Cthrc1-null mice and increased in Cthrc1 transgenic mice, respectively. Furthermore, BrdU incorporation assays showed that Cthrc1 accelerated osteoblast proliferation in vitro and in vivo. In addition, overexpression of Cthrc1 in the transgenic mice attenuated ovariectomy-induced bone loss.Conclusions/Significance
Our results indicate that Cthrc1 increases bone mass as a positive regulator of osteoblastic bone formation and offers an anabolic approach for the treatment of osteoporosis. 相似文献160.
Rolland M Heckerman D Deng W Rousseau CM Coovadia H Bishop K Goulder PJ Walker BD Brander C Mullins JI 《PloS one》2008,3(1):e1424