Shyu Conrad, Soule Terry, Bent Stephen J, Foster James A, Forney Larry J
Department of Computer Science, University of Idaho, Moscow, ID 83844, USA.
Microb Ecol. 2007 May;53(4):562-70. doi: 10.1007/s00248-006-9106-0. Epub 2007 Apr 4.
A web-based resource, Microbial Community Analysis (MiCA), has been developed to facilitate studies on microbial community ecology that use analyses of terminal-restriction fragment length polymorphisms (T-RFLP) of 16S and 18S rRNA genes. MiCA provides an intuitive web interface to access two specialized programs and a specially formatted database of 16S ribosomal RNA sequences. The first program performs virtual polymerase chain reaction (PCR) amplification of rRNA genes and restriction of the amplicons using primer sequences and restriction enzymes chosen by the user. This program, in silico PCR and Restriction (ISPaR), uses a binary encoding of DNA sequences to rapidly scan large numbers of sequences in databases searching for primer annealing and restriction sites while permitting the user to specify the number of mismatches in primer sequences. ISPaR supports multiple digests with up to three enzymes. The number of base pairs between the 5' and 3' primers and the proximal restriction sites can be reported, printed, or exported in various formats. The second program, APLAUS, infers a plausible community structure(s) based on T-RFLP data supplied by a user. APLAUS estimates the relative abundances of populations and reports a listing of phylotypes that are consistent with the empirical data. MiCA is accessible at http://mica.ibest.uidaho.edu/.
已开发出一种基于网络的资源——微生物群落分析(MiCA),以促进利用16S和18S rRNA基因的末端限制性片段长度多态性(T-RFLP)分析进行的微生物群落生态学研究。MiCA提供了一个直观的网络界面,可访问两个专门程序以及一个格式特殊的16S核糖体RNA序列数据库。第一个程序使用用户选择的引物序列和限制性内切酶对rRNA基因进行虚拟聚合酶链反应(PCR)扩增并对扩增子进行酶切。这个程序,即计算机模拟PCR和酶切(ISPaR),使用DNA序列的二进制编码来快速扫描数据库中的大量序列,以寻找引物退火位点和酶切位点,同时允许用户指定引物序列中的错配数。ISPaR支持使用多达三种酶进行多次酶切。5'和3'引物与近端酶切位点之间的碱基对数可以以各种格式报告、打印或导出。第二个程序APLAUS根据用户提供的T-RFLP数据推断出一个合理的群落结构。APLAUS估计种群的相对丰度,并报告与经验数据一致的系统发育型列表。可通过http://mica.ibest.uidaho.edu/访问MiCA。