多变鱼腥藻藻胆体的分离和荧光鉴定其完整性与解离程度

ISOLATION OF PHYCOBILISOMES OF ANABAENA VARIABILIS AND FLUORESCENCE IDENTIFICATION OF THEIR INTACTNESS AND DISSOCIATION

  • 摘要: 完整藻胆体的室温荧光峰位于678nm附近,而不完整藻胆体其峰位于673nm以下。在液氮温度下,完整藻胆体的F686与F666相对荧光强度比值超过10,F686与F655之比值超过20。不完整藻胆体的F686与F666和F686与F655之比值远低于完整藻胆体。可用室温荧光峰的波长位置和液氮温度下F686与F655和F666的相对荧光强度比值来判断藻胆体的完整性和解离程度。而液氮温度下F686与F655,F666之比值是更灵敏的指标。

     

    Abstract: Phycobilisomes were isolated from Anabacna variabilis. The cells were disrupted by sonication, treated with 2% Triton X-100 and ultracentrifuged on a sucrose step gradients. Isolation was carried out in 0.9 M Na-K-Phosphate buffer(pH 7.0) at 20℃. Phycobilisomes were recovered in the dark blue band.There are three liquid nitrogen temperature fluorescence emission bands in phyco bilisomes of A. variabilis. In intact phycobilisomes, the relative fluorescence of 686nm was very high, those of 655nm and 666nm were low, the ratios of relative fluorescence of 686nm to 655nm was more than 20, and that of 686nm to 666nm was more than 10. In non-intact phyeobilisomes, the relative fluorescence of 686 nm deereased, those of 655nm and 666mn increased. The room temperature fluorescence emission maximum of intact phycobilisomes was Iocated around 678nm, that of non-intact phycobilisomes was less than 673nm.We suggest that the main criteria to be used in assessing intact phycobilisomes of A. variabilis should be the fluorescence emission around 678nm at room temperature and the ratios of relative fluorescence of 686nm to 655nm and 666nm at liquid nitrogen temperature.Liquid nitrogen temperature fluorescence excitation spectra of F655, F666 and F68 were studied.

     

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