菜青虫细胞体外诱导产生抗菌肽初报

The report of inducible antibacterial peptides using cultured Pieris rapae insect cells

利用灭活的大肠杆菌诱导昆虫离体细胞Pr-49和Tn-5B1细胞,对诱导产物进行抑菌试验,结果显示形成了明显的抑菌圈,Tricine-SDS-PAGE分析,Pr-49细胞诱导后多出了1条分子量约7 000新的蛋白带,而且至少有3条加强蛋白带.诱导Tn-5B1细胞发现多出2条蛋白带,其中1条分子量为已证实的7 691,通过直接取浓缩上清诱导物对比浓缩上清和细胞混合物的电泳分析,新的蛋白条带为分泌型物质.初步断定离体培养的Pr-49细胞和Tn-5B1细胞经灭活的大肠杆菌诱导分泌了具有抗菌活性的物质.

Cultured Pr-49 and Tn-5B1 insect cell lines were induced with heat-killed Esherichia coli DH5α cells. Analysis by bacterial growth inhibition experiments and tricine-SDS-PAGE of supernatant from induced cells indicated that they both generated obvious inhibition zones and novel peptide bands compare to control. Induced Pr-49 cells produced a novel band with a molecular of under 7kd and three bands of higher MW. Two bands from induced Tn-5B1 cells one of which MW has been proved to be 7691D. Analysis by tricine-SDS-PAGE of both supernatant and mixture of induced Tn-5B1 cell lines showed that the two bands are secreted in the medium. Taken together our results showed that antibacterial substances were produced and Secreted into media from induced insect cell lines.