摘要：Curcin, a ribosome-inactivating protein with a molecular weight of about 28.2 kD, which strongly inhibits the protein synthesis in rabbit reticulocyte lysate system with an IC50 value of about (0.19 +/- 0.01) nmol/L, was purified from the seeds of Jatropha curcas L. The protein has the activity of rRNA N-glycosidase. Degenerate primers were designed based on the N-terminal partial sequence from purified curcin. The full-length curcin cDNA by RT-PCR and 5'-RACE was cloned. The deduced amino acids sequence indicates that a preprotein with 20 amino acid residues is first translated and then processed to a mature protein with 251 amino acids. The deduced amino acids sequence shares homology of 33% and 57% to those of type I ribosome-inactivating proteins (RIPs) and A chain of type II RIPs, respectively. The sequence encoding mature curcin was integrated into the pQE-30 vector for expression in Escherichia coli strain M15 (pREP4). The purified recombinant curcin was able to inhibit protein synthesis in rabbit reticulocyte lysate system.

摘要：[Objective] This study aimed to clone and analyze the sequence of CHS gene from Acer truncatum leaves. [Method] Using A. truncatum cultivars No.1-6 as experimental materials, total RNA was extracted from A. truncatum leaves with the modified CTAB method. CHS gene sequences were downloaded from the NCBI and aligned by BLAST. Degenerate primers were designed by DNAMAN and Primer- premier5 to amplify the target band. CHS gene fragment was amplified by RT-PCR and ligated to pMD18-T vector. The identified positive colonies were sequenced. [Result] A 1 365 bp fragment was amplified. Sequence analysis suggested that the obtained fragment encoded 365 amino acids and shared above 90% homology to nucleotide sequence of CHS gene from A. palmatum and A. [Conclusion] In this study, CHS gene was successfully cloned from A. truncatum for the first time, which laid the foundation for efficient utilization of CHS gene.

摘要：[Objective]The aim was to explore the function of WRKY transcription factor in tomato.[Method]The primers were designed in this study according to the obtained WRKY fragments,and the total RNA from tomato treated with 100 μmol/L of JA for 6 h was used as the template for RT-PCR.[Result]The 608 bp fragment was obtained from tomato with RT-PCR *** analysis indicated that this sequence contained WRKYGQK conservative domain and the similarity with Capsicum annuum WRKY-c and Nicotiana tabacum NtWRKY-7 were 79% and 74%,respectively.[Conclusion]WRKY gene sequence in tomato was cloned successfully.

摘要：[Objective] The aim was to clone the conserved sequences of cry gene and express them in Rosetta （DE3）. [Method] Specific primers were designed according to NCBI database information and the conserved sequences of cry gene were amplified by PCR from Bt transgenic cotton. Then recombinant plasmids were constructed and expressed in E. coil strain Rosetta （DE3）. Finally, the effects of different concentrations and inducing time of IPTG on the expression level of protein were investigated. [Result] Two conserved sequences （304 and 853 bp respectively） of cry gene were amplified. The result of SDS-PAGE confirmed that the recombinant plasmids pGEX-4t-I-304 and pGEX-4t-1-853 could express fusion proteins by IPTG induction and the molecular weight of protein products was 39 and 62.4 kDa respectively, which was in accordance with predicted result. The optimal protein ex- pression conditions were confirmed as induction with 0.15 mmol/L IPTG for 7 h. [Conclusion] This study prepared the ground for the further detection of Bt transgenic crops.

摘要：[Objective] The aim was to obtain the full-length cDNA sequence of Dunaliella parva β-actin gene.[Method] Based on the highly conserved amino acid regions of known β-actin,a pair of degenerate primers was synthesized to amplify 533 bp cDNA sequences in Dunaliella ***,the 5＇ genomic DNA and 3＇ cDNA sequences were obtained by Genome walking and 3＇-RACE technology based on the obtained *** to the sequences of the 5＇-termini and 3＇-termini,specific primers were synthesized to obtain the full-length cDNA.[Result] The full-length β-actin cDNA included 1 137 bp open reading frame（ORF）,617 bp of 3＇ noncoding *** analysis indicated that the highest similarity was found between Dunaliella parva and Dunaliella *** Dunaliella parva β-actin also showed wide similarity with other algae.[Conclusion] The full-length cDNA sequence of *** was firstly obtained,which was highly conserved.

摘要：[Objective] This study aimed to clone and analyze the cDNA of ATPase β subunit gene from Eleutherococcus senticosus.[Method] A pair of homologous primers was designed according to the chloroplast ATPase β subunit gene sequences of the known species;then the gene cDNA of *** were amplified by RT-PCR and compared with that of the known species;its structure was predicted finally.[Result] 1 099 bp of ATPase beta subunit cDNA of *** which encodes 366 amino acids was amplified by *** comparison and structure prediction showed that amino acids encoded by the ATPase beta subunit gene of *** shared the highest homology,up to 96.41% with that of Oryza *** the secondary structure,the protein contained 171 alpha helixes accounting for 46.72%,53 extended strands accounting for 14.48%,27 beta sheets accounting for 7.38% and 115 random coils which took up 31.42%.The amino acids 262-271 were the symbolic site of ATPase β *** whole peptide chain had no obvious hydrophobic region and was primarily confirmed as a hydrophilic protein.[Conclusion] The cNDA of ATPase β subunit gene cloned from *** in this study will provide reference for learning the effect of energy metabolism on secondary metabolism,structure and function of ATPase in ***.

摘要：Ethylene plays an extensive role in plant growth and development.. 1-aminocyclopropane-1-carboxylate （ACC） oxidase （ACO） is the key enzyme in ethylene biosynthesis. In this study, a 354 g DNA and a 213 bp cDNA base pair （bp） candidate fragment was amplified from pepper with primers derived from the ACO sequence （AJ011109） reported by Ernesto. The putative new gene was analyzed by bioinformatics tools.

摘要：基因工程是目前国内外十分活跃的研究领域,它的特点是按照人们预先设计的蓝图,把某一目的基因的DNA和载体DNA连接起来,把它引入细菌或酵母菌(也可切除原有的具有不良作用的基因),得到一重组微生物,即工程菌。再由工程菌产生这一目的基因的产物,即在分子水平上操作,细胞水平上表达。因此,基因工程也称之为基因克隆(gene cloning)和重组DNA(recombinant DNA)技术。