Decreasing erucic acid level by RNAi-mediated silencing of fatty acid elongase 1 (BnFAE1.1) in rapeseeds (Brassica napus L.)

The β-ketoacyl CoA synthase encoded by fatty acid elongase 1 gene (BnFAE1.1) is a rate-limiting enzyme regulating biosynthesis of erucic acid in rapeseeds (Brassica napus). To develop low level of erucic acid in rapeseeds by intron-spliced hairpin RNA, an inverted repeat unit of a partial BnFAE1.1 gene interrupted by a spliceable intron was cloned into pCAMBIA3301, and a seed-specific (Napin) promoter was used to control the transcription of the transgene. Four transgenic plants harboring a single copy of transgene were generated. Expression of endogenous BnFAE1.1 gene in developing T3 seeds was significantly reduced. In mature T3 seeds, erucic acid was decreased by 60.8 to 99.1% compared with wild type seeds, and accounted for 0.36 to 15.56% of total fatty acids. The level of eicosenoic acid was also greatly decreased. Furthermore, it resulted in a significant increase in the level of oleic acid, but total fatty acid content in T3 seeds was the same with that in wild type seeds. In conclusion, the expression of endogenous BnFAE1.1 was efficiently silenced by the designed RNAi silencer, causing a significant down-regulation in the level of erucic acid. Therefore, the RNAi-mediated post-transcriptional silencing of FAE1 gene to reduce oleic acid in rapeseeds was an efficient method to breed some new B. napus lines.Key words: Brassica napus L., fatty acid elongase, intron-spliced hairpin RNA, down-regulation, erucic acid

Identification, cDNA cloning, and targeted deletion of p70, a novel, ubiquitously expressed SH3 domain-containing protein

In a screen for proteins that interact with Jak2, we identified a previously uncharacterized 70-kDa protein and cloned the corresponding cDNA. The predicated sequence indicates that p70 contains an SH3 domain and a C-terminal domain with similarities to the catalytic motif of phosphoglycerate mutase. p70 transcripts were found in all tissues examined. Similarly, when an antibody raised against a C-terminal peptide to analyze p70 protein expression was used, all murine tissues examined were found to express p70. To investigate the in vivo role of p70, we generated a p70-deficient mouse strain. Mice lacking p70 are viable, develop normally, and do not display any obvious abnormalities. No differences were detected in various hematological parameters, including bone marrow colony-forming ability, in response to cytokines that utilize Jak2. In addition, no impairment in B- and T-cell development and proliferative ability was detected

Beta-type Ti-Nb-Zr-Cr alloys with large plasticity and significant strain hardening

A series of Ti-25Nb-8Zr-xCr (x = 0, 2, 4, 6, 8 wt%) alloys were designed based on DV-Xα cluster method and e=a-Δr diagram with an anticipation to obtain high plasticity and significant strain hardening. The designed alloys were produced through cold crucible levitation melting technique in order to effectively investigate their micro-structures and mechanical properties. The addition of Cr significantly enhances the β stability in the microstructures of the Ti-25Nb-8Zr-xCr alloys. Both yield strength and hardness of the studied alloys increase due to the effect of solid-solution strengthening. By contrast, the plasticity, maximum strength and strain hardening rate are influenced by theβstability as well as the distinct deformation mechanisms. None of the alloys comprising Cr fail up to 100 kN (the load capacity used) and all show impressive plasticity (~75%) and superior maximum compressive strength (~4.5 GPa) at 100 kN. Moreover, the deformation bands, which are found around the hardness indentations, are analyzed for all the investigated alloys. The fracture behaviors of the Ti-25Nb-8Zr-xCr alloys are also studied to observe the characteristics related to crack propagation, plastic deformation and the formation of shear bands