中国医学创新
中國醫學創新
중국의학창신
MEDICAL INNOVATION OF CHINA
2014年
36期
13-15
,共3页
徐春华%刘越%肖利民%邓圣泽%李东海
徐春華%劉越%肖利民%鄧聖澤%李東海
서춘화%류월%초이민%산골택%리동해
TK基因%IL-18基因%腺病毒载体%构建%鉴定
TK基因%IL-18基因%腺病毒載體%構建%鑒定
TK기인%IL-18기인%선병독재체%구건%감정
TK gene%IL-18 gene%Adenovirus vctor%Construction%Identification
目的:分别构建含TK及IL-18基因的重组腺病毒载体,并进行鉴定。方法:通过DNA重组技术,构建含有目的基因的复制缺陷型腺病毒AdCMV-TK及AdCMV-IL-18;将上述重组腺病毒液感染293细胞,提取DNA,进行PCR鉴定;将病毒液感染293细胞,扩增病毒,用OD260法测定病毒滴度。结果:经PCR鉴定,鉴定正确的腺病毒命名为AdCMV-TK及AdCMV-IL-18,测定病毒滴度AdCMV-TK=1.28×109 PFU/mL;AdCMV-IL-18=1.28×109 PFU/mL。结论:成功构建了含HSV-TK基因及IL-18基因重组腺病毒载体,为肿瘤的基因治疗提供了理论依据及实验数据。
目的:分彆構建含TK及IL-18基因的重組腺病毒載體,併進行鑒定。方法:通過DNA重組技術,構建含有目的基因的複製缺陷型腺病毒AdCMV-TK及AdCMV-IL-18;將上述重組腺病毒液感染293細胞,提取DNA,進行PCR鑒定;將病毒液感染293細胞,擴增病毒,用OD260法測定病毒滴度。結果:經PCR鑒定,鑒定正確的腺病毒命名為AdCMV-TK及AdCMV-IL-18,測定病毒滴度AdCMV-TK=1.28×109 PFU/mL;AdCMV-IL-18=1.28×109 PFU/mL。結論:成功構建瞭含HSV-TK基因及IL-18基因重組腺病毒載體,為腫瘤的基因治療提供瞭理論依據及實驗數據。
목적:분별구건함TK급IL-18기인적중조선병독재체,병진행감정。방법:통과DNA중조기술,구건함유목적기인적복제결함형선병독AdCMV-TK급AdCMV-IL-18;장상술중조선병독액감염293세포,제취DNA,진행PCR감정;장병독액감염293세포,확증병독,용OD260법측정병독적도。결과:경PCR감정,감정정학적선병독명명위AdCMV-TK급AdCMV-IL-18,측정병독적도AdCMV-TK=1.28×109 PFU/mL;AdCMV-IL-18=1.28×109 PFU/mL。결론:성공구건료함HSV-TK기인급IL-18기인중조선병독재체,위종류적기인치료제공료이론의거급실험수거。
Objective:To construct and identify the recombinant adenovirus vector containing TK gene and IL-18 gene resepectively.Method: Replication-deficient adenovirus vector containing AdCMV-TK gene and AdCMV-IL-18 gene were constructed by using DNA recombination;293 cells were transduced by AdCMV-TK gene and AdCMV-IL-18 gene,then DNA of those transduced cells was extracted and PCR was conducted; AdCMV-TK and AdCMV-IL-18 recombinant adenovirus were efficently amplified in 293 cells and the titer of the virus was determined by OD260 assay.Result:the AdCMV-TK gene and AdCMV-IL-18 gene were identified correctly by PCR,the titer of the virus:AdCMV-TK=1.28×109 PFU/mL;AdCMV-IL-18=1.28×109 PFU/mL.Conclusion:Replication-deficient Recombinant Adenovirus Vector Containing HSV-TK gene and IL-18 gene are constructed successfully, it would provide some basic theory and experimental data for the tumor gene therapy.