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11.
Reversal of Fv-1 host range by in vitro restriction endonuclease fragment exchange between molecular clones of N-tropic and B-tropic murine leukemia virus genomes. 总被引:10,自引:10,他引:0 下载免费PDF全文
L R Boone F E Myer D M Yang C Y Ou C K Koh L E Roberson R W Tennant W K Yang 《Journal of virology》1983,48(1):110-119
We molecularly cloned unintegrated viral DNA of the BALB/c endogenous N-tropic and B-tropic murine leukemia retroviruses and in vitro passaged N-tropic Gross (passage A) murine leukemia retroviruses. Recombinant genomes were constructed in vitro by exchanging homologous restriction enzyme fragments from N- or B-tropic parents and subsequent recloning. Infectious virus was recovered after transfection of these recombinant genomes into NIH-3T3 cells and cocultivation with the Fv-1 nonrestrictive SC-1 cells. XC plaque assays of recombinant virus progeny on Fv-ln and Fv-lb cells indicated that the Fv-l host range was determined by sequences located between the BamHI site in the p30 region of the gag gene (1.6 kilobase pairs from the left end of the map) and the HindIII site located in the pol gene (2.9 kilobase pairs from the left end of the map). 相似文献
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视网膜神经节细胞的纯化和体外存活 总被引:1,自引:0,他引:1
我们用特异性抗体Thy1.1结合尼龙筛方法分离和纯化新生大鼠视网膜神经节细胞,比较顶盖提取液对这些纯化细胞的作用。预先以快蓝(fast blue,FB)逆行标记的视网膜细胞悬液,接种在包被了Thy1.1抗体的培养皿上30分钟,冲洗未粘附的细胞,显微镜下计数粘附细胞中FB标记的视网膜神经节细胞纯度的百分比,最高为95%。用孔径15μm尼龙筛方法分离的纯度仅为60±5%。上述两种方法纯化的视网膜神经节细胞,仅在有顶盖提取液存在时,细胞存活并生长活跃,胞体大且有突起伸出。MTT微量比色法测定培养24小时纯化细胞存活的光密度(OD)值,显示以Thy1.1特异性抗体纯化的细胞,其OD值比值(+Te/-Te)是12.3(0.111/0.009);以尼龙筛纯化的OD值比值(+Te/-Te)是6.4(0.102/0.016);未经纯化的OD值比值(+Te/-Te)是3.8(0.095/0.025)。在上述三组中,加Te与无Te细胞生存的OD值比较,相差均非常显著(P<0.01)。结论:在纯化的视网膜神经节细胞的培养中,由于排除了其他细胞所引起的非特异性反应,神经节细胞能够更直接地反映顶盖提取液的生物效应;视网膜神经节细胞纯度越高,其作用越显著。 相似文献
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九种罕见的人类染色体异常核型报告 总被引:1,自引:1,他引:0
ReportonNineRareSpeciesofHumanChromosomalAbnormalKaryotypesHanWeitian;QuOu;YuPing;JiangMiao(LiaoningResearchInstituteofFamilyPlanning,Shenyang110031)自1983年以来,我们对数千例不育及胚胎丢失等生殖异常患者进行细胞遗传学研究,发现大量异常染色体核型,而且异常种类繁多,已报道世界首报人类异常核型25种[1]。最近,在不良妊娠患者中又发现9种异常核型,经湖南医科大学医学细胞遗传学国家培训中心鉴定,为世界首次报道.现报告如下。1病例摘要及核型例1男,30岁,表型及智力正常,其妻子妊娠两次,均在无任何诱因情… 相似文献
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采到骨状拟夏孢锈(Uredinopsis ossiformis Kamei)为中国新记录种。 相似文献
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Compartmentalization of SHC, GRB2 and mSOS, and hyperphosphorylation of Raf-1 by EGF but not insulin in liver parenchyma. 总被引:15,自引:1,他引:14 下载免费PDF全文
Rat liver parenchyma harbors equal numbers of epidermal growth factor (EGF) and insulin receptors. Following administration of a saturating dose of EGF (10 micrograms/100 g body weight), there was a rapid (t1/2 approximately 1.1 min) internalization of receptor coincident with its tyrosine phosphorylation at residue 1173 and receptor recruitment of the adaptor protein SHC, its tyrosine phosphorylation and its association with GRB2 and the Ras guanine nucleotide exchange factor, mSOS, largely in endosomes. This led to a cytosolic pool of a complex of tyrosine-phosphorylated SHC, GRB2 and mSOS. It was demonstrated that these constituents were linked to Ras activation by the characteristic decrease in Raf-1 mobility on SDS-PAGE, which was maintained for 60 min after a single bolus of administered EGF. While insulin administration (15 micrograms/100 g body weight) led to insulin receptor beta-subunit tyrosine phosphorylation and internalization, there was little detectable tyrosine phosphorylation of SHC, recruitment of GRB2, association of a complex with mSOS or any detectable change in the mobility of Raf-1. Therefore, in normal physiological target cells in vivo, distinct signaling pathways are realized after EGF or insulin receptor activation, with regulation of this specificity most probably occurring at the locus of the endosome. 相似文献
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The importance of spontaneous lac host-trees for preserving the fragile agroecosystem of the main lac producing areas of China, located in the tropical part of Yunnan province, is analyzed. In upland fields, the fire-resistant vegetation consists mainly of these trees. Farmers deliberately preserve young sprouts during the cultivation cycle. This benefits both soil fertility and lac production. Host-trees, mainly Dalbergia and Ficus, form a range of multipurpose trees which although limited to some ten species allows maximum use of the heterogeneities of the village territory. Finally, lac host-trees are associated with some environmental values important to farmers. This circumstance contributes to the sustainability of lac host-tree agroforestry in spite of current problems in the lac market. 相似文献
19.
Expression and characterization of virus-like particles containing rubella virus structural proteins. 总被引:7,自引:1,他引:6 下载免费PDF全文
Rubella virus (RV) virions contain two envelope glycoproteins (E1 and E2) and a capsid protein (C). Noninfectious RV-like particles (VLPs) containing three structural proteins were expressed in a BHK cell line (BHK-24S) by using an inducible promoter. These VLPs were found to resemble RV virons in terms of their size, their morphology, and some biological activities. In immunoblotting studies, VLPs were found to bind similarly to native RV virions with 10 of a panel of 12 RV-specific murine monoclonal antibodies. Immunization of mice with VLPs induced specific antibody responses against RV structural proteins as well as virus-neutralizing and hemagglutination-inhibiting antibodies. After immunization of mice with VLPs, in vitro challenge of isolated lymphocytes with inactivated RV and individual RV structural proteins stimulated proliferation. Our data suggest the possibility of using VLPs as immunogens for serodiagnostic assays and RV vaccines. 相似文献
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The influence of temperature, shoot age, and medium on gall induction by Subanguina picridis on Russian knapweed (Acroptilon repens) was examined in vitro. The optimal temperature for gall formation was 20 C. Gall induction was delayed as the temperature decreased, and decreased as shoot age increased. Bud primordia (0-day-old shoots and 5-day-old shoots) with an average length of 4.2 mm and 7.9 mm were the most suitable tissues for nematode development and gall formation. Gall formation was more effective on B5G medium than on MSG. Young shoots under slow growth were most suitable for mass rearing of S. picridis. 相似文献