使用抗生素是分子生物学实验常用方法,有许多筛选实验都需要用到抗生素。多数情况下,这里提到的抗生素是用来筛选细菌/真菌的,并非像医生一般,使用抗生素来治疗病人病症。BioEngX小编想在本文简单罗列各种实验室使用到的抗生素,介绍各种抗生素的适用范围和建议浓度。在文末有一个索引表格,方便对照,有需要的就把它下载取走吧~
在正文开始之前先介绍几个用在本文的缩写
- Gm+=Gram positive/革兰氏阳性细菌
- Gm-=Gram negative/革兰氏阴性细菌
- My=Mycoplasma/支原体
- Ye=Yeast/酵母菌
- Fu=Fungus/真菌
Ampicillin/氨苄青霉素
氨苄青霉素是一种β-內酰胺类抗生素(β-lactam),能够渗透格兰仕阳性及部分革兰氏阴性菌。能够和肽链转移酶(transpeptidase, a.k.a, penicillin-binding protein)进行不可逆转性结合,从而抑制细菌细胞壁的形成,最终导致细胞裂解。这便是氨苄青霉素杀死细菌的基本机制。
抗氨苄质粒是通过引入β-内酰胺酶(β-lactamase)基因,使细胞具备表达β-内酰胺酶的能力,将生存环境中的氨苄青霉素分解使其无效。
Range: Gm+/Gm-
Conc: 100-200 ug/mL
Amphotericin B/两性霉素B
两性霉素B可与固醇类物质结合,在细胞膜上形成孔洞,导致细胞内成分(氨基酸、葡萄糖……)外泄,细胞无法正常代谢,最终导致细胞裂解死亡。由于细菌细胞膜上没有固醇类分子,所以两性霉素B对细菌无抑制或杀灭作用;但真菌和动物细胞膜上都有固醇类分子,所以在使用的时候一定要注意安全。
Range: Fu/Ye/My
Conc: 2.5 ug/mL
Carbenicillin/羧苄青霉素
从名字我们也可以看出,羧苄青霉素和氨苄青霉素十相似。羧苄青霉素对革兰氏阳性细菌比氨苄青霉素稍弱,但可对抗更广的革兰氏阴性菌菌种。作用机制与氨苄青霉素一致。
Range: Gm+/Gm-
Conc: 100 ug/mL
Chloramphenicol/氯霉素
广谱抗生素,对革兰氏阳性菌有明显对抗左右。氯霉素能抑制细菌核糖体的肽酰转移酶从而抑制细菌蛋白质的合成,导致细菌死亡。现在在临床上已少用氯霉素了,主要原因是其副作用会导致不良性贫血。但是,在分子生物实验里我们还能经常见到这种抗生素。值得一提的是,在上世纪五十年代,氯霉素被发现并分离于Streptomyces venezuelae,看这个venezue…单词,可以知道,这个菌最早发现于南美洲的委内瑞拉~~
Range: Gm+/Gm-
Conc: 5-10 ug/mL
氯霉素乙酰转移(Chloramphenicol acetyltransferase,CAT)能够破坏氯霉素的杀菌特性。CAT能够催化一个化学反应,将一个乙酰基团从乙酰辅酶A转移到氯霉素,这样就会阻止氯霉素与核糖体的结合,从而使之失去效果。
Erythromycin/红霉素
大环内酯类抗生素,作用机制与氯霉素基本相似。红霉素于上世纪五十年代发现于放线菌类的Saccharopolyspora erythraea 并纯化出来。
Range: Gm+/My
Conc: 100 ug/mL
Kanamycin/卡那霉素
氨基糖酐类抗生素,作用机理也是阻止细菌蛋白质的形成,但与氯霉素或红霉素的具体机制不一样,它阻碍Met-tRNA与核糖体复合物的结合,干扰的是蛋白质合成的起始步骤。卡那霉素最早分离于链霉菌群中的Streptomyces kanamyceticus。
Range: Gm+/Gm-/My
Conc: 100 ug/mL
Gentamycin/庆大霉素
与卡那霉素同属一类抗生素,作用机制相似。
Range: Gm+/Gm-
Conc: 25-50 ug/mL
Neomycin/新霉素
与卡那霉素、庆大霉素同属一类,作用机制也相似。
Range: Gm+/Gm-
Conc: 25-50 ug/mL
Nystatin/耐斯菌素
抗真菌类抗生素,最早提炼于链霉菌群中的Streptomyces noursei。作用机制与两性霉素B相似,也是导致细胞膜孔洞形成,胞内物质外泄,最终细胞死亡。
Range: Ye/Fu
Conc: 50 ug/mL
Rifampicin/甲哌利福霉素/利福平
广谱抗生素。作用机制是抑制细菌RNA多聚酶的形成,从而阻止细菌蛋白的合成。但值得注意的是,T7 RNA多聚酶的形成不会受到影响,所以常用于筛选T7起始子引导的蛋白表达。
Range: Gm+/Gm-
Conc: 50 ug/mL
Streptomycin/链霉素
第一个被发现的氨基糖苷类抗生素,第一个用于有效治疗肺结核的抗生素,当然这些事情都发生在70年前。发现者瓦克斯曼因此获得1952年的诺贝尔奖。作用机制也是与核糖体结合,干扰蛋白合成,导致细菌死亡。
Range: Gm+/Gm-
Conc: 50 ug/mL
Tetracyclin/四环霉素
聚酮类广谱抗生素,很多链霉菌都可生产这类抗生素。杀菌机制也是干扰蛋白合成。
Range: Gm+/Gm-
Conc: 50 ug/mL
索引图如下所示:
还有什么抗生素是实验室常用却没有在文章中提到的,请在文末留言。
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