Olivares-Fuster O, Arias C R
Department of Fisheries and Allied Aquacultures, Auburn University, Auburn, AL 36849, USA.
Lett Appl Microbiol. 2008 Jun;46(6):605-12. doi: 10.1111/j.1472-765X.2008.02366.x. Epub 2008 Apr 25.
To identify specific sequences in the fish pathogen Flavobacterium columnare not shared by Flavobacterium johnsoniae.
Suppressive subtractive hybridization (SSH) was used to selectively amplify and clone F. columnare-specific sequences. A highly virulent strain of F. columnare was used as tester and the type strain of F. johnsoniae was used as driver. After library construction, 192 clones were selected and sequenced. From those, 110 clones contained unique F. columnare-specific sequences that were verified using dot blot hybridization. Sequence sizes ranged from 55 to 872 bp with 45,363 bp sequenced in total.
Specific F. columnare sequences representing all but one (motility related) functional categories were annotated. Several putative virulence factors were identified in F. columnare such as a collagenase, a chondroitinase, proteases, as well as drug resistance and iron transport-related genes.
Suppressive subtractive hybridization is a cost-effective method for identifying genetic differences between Flavobacterium spp. The number of sequences available from F. columnare has been doubled.
鉴定柱状黄杆菌中约翰逊氏黄杆菌所没有的特定序列。
采用抑制性消减杂交（SSH）技术选择性扩增并克隆柱状黄杆菌特异性序列。以一株高毒力柱状黄杆菌作为测试菌株，约翰逊氏黄杆菌的模式菌株作为驱动菌株。构建文库后，挑选192个克隆进行测序。其中，110个克隆含有独特的柱状黄杆菌特异性序列，通过斑点杂交进行了验证。序列大小在55至872 bp之间，总共测序45,363 bp。
除一个（与运动性相关）功能类别外，代表所有其他功能类别的柱状黄杆菌特异性序列均已注释。在柱状黄杆菌中鉴定出了几种假定的毒力因子，如胶原酶、软骨素酶、蛋白酶以及耐药性和铁转运相关基因。
抑制性消减杂交是一种鉴定黄杆菌属之间遗传差异的经济有效方法。柱状黄杆菌可用序列数量增加了一倍。
