摘要
目的研究巨噬细胞对新生隐球菌B3501标准株的主要毒性基因表达影响。方法将对数生长期的J774.16巨噬细胞分别与新生隐球菌野生株B3501共孵育4h,收集被J774.16吞噬的B3501作为实验组,提取实验组和在37℃条件下5%二氧化碳单独培养的对照组B3501的RNA,采用实时荧光定量PCR技术,检测J774.16细胞内和对照组B3501的CNLAC1、CAP60、URE1、NMT表达的差异。结果实验组中新生隐球菌的CAP6,CNLAC1,NMT及URE1基因的mRNA在每百万看家基因(GAPDH基因)中的平均含量分别为(2.698±0.084)×104,(1.806±0.322)×103,(2.267±0.074)×104和(4.041±0.271)×104;而对照组这四种毒性因子基因含量分别为:(1.139±0.183)×106,(9.324±5.028)×103,(1.326±0.028)×106和(1.307±0.001)×106,均远比实验组高,其中以NMT最为明显。结论新生隐球菌被巨噬细胞吞噬后,主要的毒性因子基因表达下降,其中以NMT最为明显,而CNLAC1下降幅度最小。
Objective To investigate the difference of main virulence genes mRNA level in Cryptococcus neoformans wild strain B3501 after phagocytosis by macrophage J774. 16. Method Co-cultured the logarithmic growing J774. 16 with Cryptococcus noeformans wild strain B3501. Collected the macrophage J774.16 challenged by yeast at co-cultured 4 hours and control group （ solely cultured at 37℃ ,5% CO2） yeast. Then their total RNA was extracted for screening the difference of main virulence factor gene CNLAC1 , CAP60, URE and NMT mRNA expression by real-time PCR. Results The production of the four virulence factor genes in the test group （ ingested yeast） （ CAP60, CNLAC 1, NMT and URE 1 ） （2. 698 ＋ 0. 084）×10^4, （ 1. 806 ± 0. 322） ×10^3, （2. 267 ± 0. 074） ×10^4 and （4.041 ±0.271 ） ×10^4 per million house keeper gene GAPDH, respectively. And those of the control group were （ 1. 139 ± 0.183 ）×10^6, （9. 324 ± 5. 028 ）×10^3, （ 1. 326 ＋ 0. 028 ） ×10^6 and （ 1. 307 ± 0. 001 ） ×10^6, respectively. The production of the four genes of the ingested Cryptococcus neoformans strain B3501 were 5-58 times less than those of the control group. Conclu- sion The four main virulence factor gene all expressed decreasingly after Cryptococcus neoformans was ingested by the maerophage. Among them, changement of N-myristoyltransferase gene （NMT） was the most significantly.
出处
《中国真菌学杂志》
2008年第6期325-328,共4页
Chinese Journal of Mycology
