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作者在经亲和色谱和反相高效液相色谱分离纯化,获得山羊单纯疱疹病毒Ⅰ型(HSV-1)特异转移因子单一活性成分(PTFC)。用高效凝胶色谱确定PTFC的分子量(Mr),在薄层色谱观察其254nm的紫外吸收点和366 nm与荧光胺结合产生的荧光点,并检测PTFC的氨基酸组成和序列.结果显示,该成分的Mr为6000;薄层色谱后出现-254 nm紫外吸收点和荧光胺显色点,两点重叠于薄层板同一位置;其HC1水解产物含有至少12种氨基酸,其中天冬氨酸含量占12种氨基酸的44.2％,经Edman降解未检测到PTFC的氨基酸序列.实验结果表明,PTFC是Mr为6000的多肽与核苷酸样物质的复合物,其多肽部分富含天冬氨酸,肽链N末端可能封闭。
The authors isolated and purified HSV-1 specific transfer factor (PTFC) by affinity chromatography and reversed-phase high performance liquid chromatography. The molecular weight (PT) of PTFC was determined by high performance gel permeation chromatography (GPC), the UV absorption of 254nm was observed by thin layer chromatography and the fluorescence spot was formed by the combination of 366 nm with fluorescamine, and the amino acid composition and sequence of PTFC were determined. Of Mr 6000; thin layer chromatography appeared -254 nm UV absorption point and fluorescent amine color point, two overlapping in the same position on the TLC plate; HC1 hydrolyzate contains at least 12 kinds of amino acids, of which aspartic acid content 44.2% of the 12 amino acids were not detected by Edman degradation of PTFC amino acid sequence.The experimental results show that PTFC is Mr 6000 complex of the polypeptide and the nucleotide-like material, the polypeptide portion is rich in aspartic acid, peptide N-terminal chain may be closed.