单纯疱疹病毒Ⅰ型(HSV—1)基因突变株的生物学特性初步研究

应用病毒空斑滴定法及动物LD_(50)法对HSV-1基因插入突变株[HSV-1(RC)]的生物学特性进行了初步研究。发现HSV-1(RC)株在不同的6种细胞系上能够生长繁植,但在BHK-21、Hep-2细胞上较其亲代HSV-1(Δ305)及HSV-1(F)株生长缓慢,且空斑形成单位(pFU)数明显低于亲代株。动物LD_(50)实验结果HSV-1(F)株LD_(50)值为2.9×10~4pFU/0.1ml,而HSV-1(6305)及HSV-1(RC)LD_(50)值为0。实验结果提示:HSV-1基因组BamHI C片段插入突变处不是该病毒在一些细胞培养上生长所必需的基因片段,但对在某些细胞系上的生长仍具有影响作用。HSV-1的TK基因和BamHI C片段插入突变处则是病毒在动物整体上毒力强度的必需基因。在HSV-1基因组BamHI C片段处插入9.7kb大小的外源基因并不影响病毒在细胞上的组装和繁殖,但其毒力降低,建立的实用空斑滴定技术适用于病毒学研究,有推广应用价值。

STUDY ON THE BIOLOGICAl CHARACTERISTICS OF HERPES SIMPLEX VIRUS TYP?1 (HSV-1) GENE MUTANT

Virus plaque and median lethal dose (LD_(50)) areused to study the biological characteristics of HSV--1insertional genc mutant [HSV--1 (RC)] HSV--1(RC) can grow on BHK--21, Hep--2, RK--13, VeroRSC and 143 cell cultures, but both its growing specdand the plaquc formation unit(pFU) are lower thanthe parent strains on BHK--21 Hep--2 LD_(50) ofHSV--1 (F) strain is 29×10~4 pFU/0.1ml, whileLD_(50) of HSV--1 (RC) and HSV--1 (△305) is 0These results indicate The site of insertrional muta-tion in HSV--1 BamHI C fragment is not the necessa-ry gene for virus growing on some cell cultures, but iteserts some influence on the virus growing on BHK--21,Hep--2. Insertional 9.7 kb alien gene in BamHI Cfragment has no influcnce on virus assemble and multipli-cation but the toxicity is lower HSV--1 TK gene andthe site of insertional mutaion in BamHI C fragment arethe necessary genes for virus toxicity on animal Thevirus plaque titer technique established is suitable tovirology studying and it has the value of being spread andapplied.