Cloning and expression of the tumstatin active peptides-T(7) and its derivant-T(7)-NGR.

To enhance the role targeting, design to link NGR sequence with tumstatin active peptides-T(7)'s C-terminal, the derivant called T(7)-NGR. The cloning vector pMD-T(7) and pMD-T(7) N were constructed by PCR and gene synthesis methods, respectively, identified by digestion and DNA sequencing. After the digested plasmids were isolated by the low melting point agarose electrophoresis, the target-fragment was cut off and mixed with the recovery of the digested vector pET28a.

[Enzyme-amplified time-resolved fluorescence detection for nucleic acid hybridization assays].

Objective: To develop a new nonisotopic detection method of enzyme-amplified time-resolved fluorescence (EATRF) or enzyme-amplified lanthanide luminescence (EALL) for nucleic acid hybridization assays, which can be applied extensively in clinical diagnosis.

Methods: The method combines the high affinity of biotin-streptavidin system, amplification of enzyme, and inherent advantage of lanthanide chelate with the background elimination of time-resolved fluorescence detection. The conversion of 5-fluorosalicyl phosphate to 5-fluorosalicylic acid (5-FSA) by alkaline phosphatase.