Isolation and Characterization of NBS-LRR Class Resistance Homologous Gene from Wheat

One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide binding site （NBS） conserved domain.Based on the RGA-CIN14,a full-length cDNA,CIN14,which was 2 987 bp encoding 880 amino acids,was obtained by using the method of the rapid amplification cDNA ends （RACE）.Bioinformatics analysis showed that the deduced amino acids of CIN14 protein consisted of a NB-ARC conserved domain and many leucine-rich repeats （LRR） domains.The phylogenetic tree analysis indicated a considerable identity of the protein encoded by CIN14 with that of wheat leaf rust resistance gene Lr1,but a lower similarity with Lr21.The expression profile of the CIN14 gene detected by semi-quantitative RT-PCR showed that the CIN14 gene was not induced by Puccinia triticina and it was a constitutive gene with low abundance in the wheat leaf tissue.The resistance homology sequence was successfully obtained,which provides the shortcut for cloning of the resistance gene in TcLr19 wheat.

Cloning and Identification of Porcine HSPC117 Gene Differentially Expressed in F_(1)Crossbreds and Their Parents

To investigate the molecular basis of porcine heterosis, suppression subtractive hybridization （SSH） was performed to detect the differences in gene expression between porcine longissimus dorsi of Meishan X Large White （MS × LW） F1 hybrids and their parents Meishan pigs. An expression sequence tag （EST） differentially expressed was found, designated as ML556, which was homologous to a hypothetical protein HSPC117, from human hematopoietic stem/progenitor cells （HSPCs）, and the full-length cDNA of porcine HSPC117 was cloned using rapid amplification of cDNA ends （RACE） method. Translation of the mRNA transcript revealed an open reading frame （ORF） of 505 amino acid residues encoding a peroxisomal targeting signal （PTS） with theoretical molecular weight of 55 kDa. Alignment analysis revealed that the deduced protein sequence exhibit 98, 98, 98, 97, and 97% identity with that of cattle, human, dog, rat, and mouse, respectively. The tissue expression analysis indicated that the porcine HSPC117 gene is highly expressed in muscle, spleen, lung, kidney, uterus, ovary and testis, moderately expressed in fat, heart, and liver, and not expressed in stomach and small intestine. The possible role of porcine HSPC117 and its relationship with porcine heterosis were discussed.