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21.
22.
免疫毒素Luffin B-Ng76对人黑色素瘤细胞的体外抑制作用 总被引:7,自引:0,他引:7
用BlueSepharoseCL-6B凝胶亲和层析法从丝瓜籽中分离纯化了单链致核糖体失活蛋白(ribosomeinac-tivatingprotein,RIP)——lufinB。并将lufinB与抗人黑色素瘤细胞单抗Ng76制成了免疫毒素,命名为lufinB-Ng76,它对体外培养的黑色素瘤细胞M21有很强的抑制作用,IC50为2.5×10-11mol/L,毒性比游离的lufinB提高4000倍,而它对非靶的HeLa细胞的毒性较M21细胞低1200倍。结果提示lufinB用于制备免疫毒素具有良好的应用前景。 相似文献
23.
细胞因子Midkine(简称MK)是新发现的一类肝素结合因子家族中的一员。1988年,Kadamatsu等利用差异杂交法在经维甲酸诱导分化的小鼠畸胎瘤细胞株HM-1中首先克隆到小鼠MK基因。人MK基因则最早是从λgt10人胚肾(20-24周)cDNA库和EMBL-3人胎盘基因组库获得。成熟 相似文献
24.
三孢布拉氏霉发酵生产β-胡萝卜素的研究 总被引:5,自引:0,他引:5
利用三孢布拉氏霉发酵生产天然β-胡萝卜素。在30L发酵罐中,平均生物量31.3g干菌重/L发酵液和胡萝卜素1213.1mp/L;在3M3发酵罐中,平均生物量38.0g干菌重/L发酵液和胡萝卜素1146.5mg/L。将中试样品经HPLC分析,在总色素中β-胡萝卜素占92~96%,其他类胡萝卜素占8~4%;在β-胡萝卜素中,反式异构体占90~95%,9-、13-、15-顺式异构体占10~5%。结晶β-胡萝卜素呈多种形状,但大多数为两端锥形的棱柱体。在胡萝卜素提取中,工艺和技 相似文献
25.
Jun Mi Yang Feng Jie Wen Yiqun Su Lin Xu Tingjian Zu Chang Liu David E. Fisher Xunwei Wu 《Pigment cell & melanoma research》2020,33(1):16-29
Primary melanocytes isolated from skin and expanded in culture have been widely used for laboratory research and clinical applications. The conventional method to isolate primary melanocytes from skin usually requires about 3–4 weeks of culture for melanocytes to grow sufficiently to passage. Considering that melanocytes comprise only 3%–7% of epidermal cells in normal human skin, it would be extremely helpful to increase the isolation efficiency and shorten the initial culture time to quickly meet various application needs. Here, we report that adding Y‐27632, a Rho kinase inhibitor, into the initial culture medium for 2 days can dramatically increase the yield of melanocytes. We found that Y‐27632 can promote keratinocyte attachment and survival in the melanocyte culture system, resulting in not only better recovery, but also increased proliferation of melanocytes by a paracrine signaling pathway. More specifically, Y‐27632 significantly induced keratinocyte expression of stem cell factor, which played an important role in enhancing the growth of melanocytes. In summary, Y‐27632 could profoundly enhance the yield of primary melanocytes in the initial culture through paracrine effects on keratinocytes. 相似文献
26.
目的探讨内生Bacillus svelezensis HBB5菌株发酵宿主植物盾叶薯蓣产薯蓣皂苷元的能力。方法接种内生B.svelezensis HBB5及B.subtilis ATCC 6633菌株(0.35×10~8 CFU/mL)至含盾叶薯蓣地下茎组织的液体培养基,32℃、165~185 r/min连续发酵108 h,检测发酵液细菌、pH、淀粉、麦芽糖、葡萄糖、淀粉酶(α-amylase)及薯蓣皂苷元溶出率等指标。结果内生B.svelezensis HBB5菌株有较强的酸、碱耐受力[pH(4.8±0.2)~(8.4±0.2)],相比B.subtilis ATCC 6633[pH(5.2±0.2)~(8.7±0.2)]差异不明显;前者达峰值生长量(60×10~(8±2) CFU/mL)明显高于后者(32×10~(8±2) CFU/mL)。发酵36~60 h时,B.svelezensis HBB5、B.subtilis ATCC 6633菌株发酵液的淀粉、麦芽糖、葡萄糖浓度达峰值,分别为(37.41±3.12)、(27.83±2.14)ng/mL,(21.06±1.25)、(16.54±1.08)ng/mL,(54.33±3.12)、(36.65±2.10)ng/mL,前者均高于后者。同时,B.svelezensis HBB5菌株维持高的α-amylase酶活性及薯蓣皂苷元溶出率。结论内生B.svelezensis HBB5菌株拥有较强的耐酸碱、降解淀粉、提高薯蓣皂苷元溶出的能力,为工业生产薯蓣皂苷元提供了一个新的方法。 相似文献
27.
Time‐resolved fluorescence as well as steady‐state absorption and fluorescence were detected in order to study the interactions between tetramethylrhodamine (TAMRA) and DNA when TAMRA was covalently labeled on single‐ and double‐stranded oligonucleotides. Fluorescence intensity quenching and lifetime changes were characterized and correlated with different DNA sequences. The results demonstrated that the photoinduced electron transfer interaction between guanosine residues and TAMRA introduced a short lifetime fluorescence component when guanosine residues were at the TAMRA‐attached terminal of the DNA sequences. The discrepancy of two‐state and three‐state models in previous studies was due to the DNA sequence selection and sensitivity of techniques used to detect the short lifetime component. The results will help the design of fluorescence‐based experiments related to a dye labeled probe. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献
28.
Shuang Zhao Lan-Tao Gou Man Zhang Li-Dong Zu Min-Min Hua Ye Hua Hui-Juan Shi Yong Li Jinsong Li Dangsheng Li En-Duo Wang Mo-Fang Liu 《Developmental cell》2013,24(1):13-25
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29.
An Xiao Zhanxiang Wang Yingying Hu Yingdan Wu Zhou Luo Zhipeng Yang Yao Zu Wenyuan Li Peng Huang Xiangjun Tong Zuoyan Zhu Shuo Lin Bo Zhang 《Nucleic acids research》2013,41(14):e141
Customized TALENs and Cas9/gRNAs have been used for targeted mutagenesis in zebrafish to induce indels into protein-coding genes. However, indels are usually not sufficient to disrupt the function of non-coding genes, gene clusters or regulatory sequences, whereas large genomic deletions or inversions are more desirable for this purpose. By injecting two pairs of TALEN mRNAs or two gRNAs together with Cas9 mRNA targeting distal DNA sites of the same chromosome, we obtained predictable genomic deletions or inversions with sizes ranging from several hundred bases to nearly 1 Mb. We have successfully achieved this type of modifications for 11 chromosomal loci by TALENs and 2 by Cas9/gRNAs with different combinations of gRNA pairs, including clusters of miRNA and protein-coding genes. Seven of eight TALEN-targeted lines transmitted the deletions and one transmitted the inversion through germ line. Our findings indicate that both TALENs and Cas9/gRNAs can be used as an efficient tool to engineer genomes to achieve large deletions or inversions, including fragments covering multiple genes and non-coding sequences. To facilitate the analyses and application of existing ZFN, TALEN and CRISPR/Cas data, we have updated our EENdb database to provide a chromosomal view of all reported engineered endonucleases targeting human and zebrafish genomes. 相似文献
30.
To study intraspecific differences in N utilization in response to enhanced UV-B radiation, field experiments were conducted on two Erigeron breviscapus populations (Huguo and Cangshan), which were respectively obtained from low altitude (UV-B sensitive) and high altitude (UBV-B resistant).The effects of soil nitrogen (N) application (0, 15, 30, 45 g m2) on free amino acid content, the activities of nitrate reductase (NR) and glutamine synthetase (GS), total nitrogen content and N mass in leaves were determined under enhanced UV-B radiation (5 kJ m2) for both populations. The results showed that under enhanced UV-B radiation: (1) increases in total N contents in leaves of the Huguo and Cangshan populations correlated with the amount of N applied. Additionally, leaf biomass of Huguo treated with 15 g m?2 N application and Cangshan with 30 g m?2 N application were higher than that of other treatments. Leaf N masses were highest in both E. breviscapus populations treated with 30 g m?2 N; (2) increases in contents of free amino acids in leaves of both E. breviscapus populations correlated with the amount of applied nitrogen; (3) increases of NR activity in leaves correlated with the amount of applied nitrogen; (4) GS activity in leaves of the Huguo and Cangshan E. breviscapus populations were highest with respective N applications of 15 g m?2 N and 30 g m?2 N. In general, under enhanced UV-B radiation, N application might affect NR and GS and change free amino acid content, resulting in changes in total nitrogen content, biomass and N mass. The optimal amount of supplemental N for N accumulation in E. breviscapus was 30 g m?2 N under enhanced UV-B radiation. 相似文献