【作者】 庄晓晟；

【导师】 梁伶；

【作者基本信息】 广西医科大学 ， 皮肤病与性病学， 2008， 硕士

【摘要】 目的初步探讨马尔尼菲青霉菌(Penicillium marneffei,PM)竹鼠寄生株和人感染株中GasC基因表达变化情况。方法随机选取15株成年竹鼠体内分离PM和15株人感染株PM在SDA液基25℃震荡培养72h。采Trizol法提取PM菌株的总RNA。荧光定量PCR技术测出两种不同来源的菌株GasC基因的表达量。统计分析两种不同来源的菌株GasC基因的表达量是否具有差异性。结果①15只成年广西野生银星竹鼠自然携带PM阳性率100%,30株菌株经在SDA液基25℃震荡培养72h后均产可溶性红色色素,但有程度差别,即色价不同。统计分析两种不同来源的PM在25℃时产色无差异(p＞0.05)。②30株菌株提取RNA并逆转录成cDNA,经PCR后电泳可见GasC基因的目的条带。③采用荧光定量PCR测出15株竹鼠寄生株PM和15株人感染株PM GasC基因的表达量,SPSS统计分析组间差异无统计学意义(p＞0.05)。④菌株的色价与GasC基因的表达量有统计学差异(p＜0.05),且色素色价越高,其菌株GasC基因表达水平就越高。结论两种不同来源的PM25℃时产色无差异;GasC基因在PM寄生株和人感染株均有表达,其表达量不存在差异,提示两组PM菌株在活力和致病力上可能不存在差异;菌株色价与GasC基因的表达相关,色素色价越高,其菌株GasC基因表达水平就越高,但分泌色素多且GasC基因表达高的菌株致病力是否较强,需进一步动物实验验证。更多还原

【Abstract】 Objectives To investigate the differential expression of the GasC gene in Guangxi Penicillium marneffei (PM) between the isolates obtained from Rhizomys pruinosus and those from Penicilliosis marneffei(PSM) patientsMethods Selected randomly 30 isolates separately obtained from Rhizomys pruinosus and those from PSM patients cultured in SDA liquid medium at 25℃after 72h. Total RNA was extracted with Trizol. Detect the expression of GasC gene in each isolates through the fluorescence quantitive PCR technic. Analysis the difference of the GasC expressed in the two group by SPSS.Results①The natural positive carrier rate of the 15 was 100 %, 30 isolates can all produced the red pigment at 25°C after 72h cultured in SDA liquid medium,and they all produced the red pigment with different level,but no difference in statistics between the two group (p>0.05) .②we can see the goal gene through the gelelectrophoresis③Detect the expression of GasC gene in each isolates through the Real Time PCR and there is not difference in statistics between the two group (p>0.05) .④The expression of GasC gene is posertivly connect with the ability for producing the pigment of PM, and the deeper the pigment shows , the higher the level of GasC gene expressed.Conclusions There is no difference in producing red pigment between the isolates obtained from Rhizomys pruinosus and those from Penicilliosis marneffei(PSM) patients; The expression of GasC gene is not difference in statistics between the two group which preliminary revealed the similarity in growth vitality and pathogenicity;The expression of GasC gene is connect with the ability for producing the pigment of PM, and the deeper the pigment shows , the higher the level of GasC gene expressed. It must be proved by the further experiment on animals whether the PM with deeper pigment and the high expression of GasC gene presents stronger pathogenicity.更多还原