Bartzatt R L, Volsky D J
Biosci Rep. 1984 Jul;4(7):551-7. doi: 10.1007/BF01121911.
Sendai virus envelopes (SVE) were isolated from Sendai virus particles by Triton X-100 solubilization and ultracentrifugation. The envelopes were reconstituted in the presence of the fluorescent dye calcein by gradual removal of the detergent with Bio-beads SM-2. The internal volume of reconstituted Sendai virus envelopes (RSVE) was determined by quenching the fluorescence of calcein with cobalt (II) ions. The internal volume of RSVE was found to be proportional to the initial SVE protein concentration in the reconstitution mixture, reaching about 18% of the total volume with 5.6 mg of SVE protein per ml. When radiolabelled cloned Epstein-Barr virus DNA fragment was included in the reconstitution mixture, the proportion of DNA associated with the vesicles much exceeded the trapping volume, indicating adsorption of DNA to the internal surface of RSVE. These determinations will allow optimization of the use of RSVE as gene-transfer vehicles.
仙台病毒包膜（SVE）通过用Triton X-100增溶和超速离心从仙台病毒颗粒中分离出来。通过用Bio-beads SM-2逐渐去除去污剂，在荧光染料钙黄绿素存在的情况下重建包膜。通过用钴（II）离子淬灭钙黄绿素的荧光来测定重建的仙台病毒包膜（RSVE）的内部体积。发现RSVE的内部体积与重建混合物中初始SVE蛋白浓度成正比，当每毫升含有5.6毫克SVE蛋白时，其内部体积达到总体积的约18%。当将放射性标记的克隆爱泼斯坦-巴尔病毒DNA片段包含在重建混合物中时，与囊泡相关的DNA比例远远超过捕获体积，表明DNA吸附到RSVE的内表面。这些测定将有助于优化RSVE作为基因传递载体的使用。
