大豆吡哆醇生物合成蛋白基因PDX的电子克隆和进化-华北农学报.pdf

华北农学报2007 , 22 ( 1) : 64- 6 8 (PDX ) 李 蕊, 孟宪萍, 胡英考, 蔡民华, 李雅轩 ( , 100037) : ( In silico cloning) EST , ( Arabidopsis thaliana) cDNA , ( Glycine max) EST , 1 280 bp ( GenB nk DQ139265) RT-PCR PCR , ( ORF, 20~ 955 bp) , 311 , , EST , : ; EST ; RT-PCR; PCR; ; : S5651 : A : 1000- 7091( 2007) 01- 0064- 05 In Silico Cloning of Pyridoxine Biosynthesis Protein Gene from Soybean and Evolution Analysis LI Rui, MENG Xi n-ping, HU Ying-k o, CAI Min-hu , LI Y-xu n ( College of Life Science, C pit l Norm l University, Beijing 100037, Chin ) Abstract: In silico cloning is developing with the project of genomic nd EST . It is new method of gene-cloning through bioinform tics.A query probe from the cDNA sequence of Arabidopsis thaliana pyridoxine biosynthesis protein w s designed to bl st Glycinemax EST d t b se ccording to the rel tive conserv tion of homologous genes nd soybe n PDX gene cDNA of 1 280 bp w s obt ined ( GenB nk Accession, DQ139265) . This sequence w s confirmed by RT- PCR, genomic PCR, molecul r cloned nd sequenced. It cont ined complete ORF, from 20 to 955 bp, encoded 311 mino cids, nd conserved with Arabidopsis thaliana, Oryza sativ , Lotus corniculatus v r. japonicus, Nicotiana tabacum, edicago truncatula nd Triticum aestivum. The results showed th t it w s n efficient technique to clone new genes by se rching EST d t b se with homologous gene of model species. Key ords: Insilico cloning; EST d t b se; RT-PCR; Genomic PCR; Glycine max; Pyridoxine biosynthesis protein ( Expressed sequence t gs, , [ 5] EST) (In silico cloning) , , B 6