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        Isolation and characterization of heat-responsive gene TaGASR1 from wheat (Triticum aestivum L.)

        Liyuan Zhang,Xiaoli Geng,Haiyan Zhang,Chunlei Zhou,Aiju Zhao,Fei Wang,Yue Zhao,Xuejun Tian,Zhaorong Hu,Mingming Xin,Yingyin Yao,Zhongfu Ni;Qixin Sun,Qixin Sun,Huiru Peng 한국식물학회 2017 Journal of Plant Biology Vol.60 No.1

        GA-stimulated transcript (GAST) family genes have been identified in numerous plant species. In this paper, we isolated and characterized a heat-responsive gene, TaGASR1, from heat tolerant variety TAM107. The complete ORF of TaGASR1 was cloned, which encoded a 98-kDa protein, and the sequence shared 51.52% similarity to OsGASR1. Analysis of the TaGASR1 promoter region showed that it contained a heat shock element (HSE) and several cis-elements involved in abiotic stress response and hormone signal transduction. Expression patterns of TaGASR1 revealed that it was strongly induced by stress factors, such as high temperature, drought, high salinity and oxidation, as well as the phytohormones, including MeJA, ACC and ABA, which suggested the TaGASR1 gene might participate in these stress and hormone signal transduction pathways. Transient expression of TaGASR1-GFP fusion proteins in onion epidermal cells indicated that TaGASR1 protein was localized to the cell membrane or cytosol. Further analysis showed that ectopic expression of TaGASR1 in Arabidopsis enhanced thermotolerance and reduced the accumulation of reactive oxygen species (ROS) after heat stress. Moreover, we also found that TaGASR1-overexpressing wheat improved tolerance to heat stress and oxidative stress.

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        Molecular Cloning, Characterization, and Expression Analysis of Chicken Δ-6 Desaturase

        Kang, Xiangtao,Bai, Yichun,Sun, Guirong,Huang, Yanqun,Chen, Qixin,Han, Ruili,Li, Guoxi,Li, Fadi Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.1

        Long-chain polyunsaturated fatty acids (LC-PUFA) promote the development of brain and vision of the fetus, relieve inflammation, inhibit oral dysplasia of rumor cell, decrease the incidence of cardiovascular disease and regulate arrhythmia. ${\Delta}-6$ desaturase is the rate-limited enzyme in the desaturation process. This study reports the cloning, characterization and tissue expression of a ${\Delta}-6$ desaturase gene in the chicken. PCR primers were designed based on the predicted sequence of chicken ${\Delta}-6$ desaturase (accession number: XM421053) and used to isolate a cDNA fragment of 1,323 bp from chicken liver. Based on the 1,323 bp fragment an EST (BI390105) was obtained by BLAST. The EST and 5'nd of the 1,323 bp fragment were partially overlapped. Gene specific primers derived from the EST were used for amplification of the 5'nd. Another gene-specific primer derived from the 1,323 bp fragment was used for amplification of the 3'nd by 3'ACE. Then the three overlapping cDNA sequences obtained were assembled with DNAMAN software and a full-length ${\Delta}-6$ desaturase of 2,153 bp was obtained. The full-length cDNA contained an ORF of 1,335 bp with a 5'ntranslated region of 147 nucleotides followed by an ATG initiation codon. Stop codon TGA was at position 1,481-1,483 bp. The deduced amino acids shared an homology above 77% with bovine, mice, orangutan, rat and human. The protein sequence had three histidine-rich regions HDFGH (HisI region), HFQHH (HisII region) and HH (HisIII region), a cytochrome $b_{5}$-like domain containing a heme-binding motif and two transmembrane domains. Sequence analysis of the chicken genomic DNA revealed that the coding sequence of chicken ${\Delta}-6$ desaturase included 12 exons and 11 introns. Semi-quantitative RT-PCR showed that the ${\Delta}-6$ desaturase expression levels were in turn liver, spleen, pancreas, lung, breast muscle, heart, and abdominal fat. The expression of ${\Delta}-6$ desaturase in liver was significantly higher than that in breast muscle (p<0.01). The expression of ${\Delta}-6$ desaturase in lung was significantly higher than that in abdominal fat (p<0.01). This is the first clone of chicken ${\Delta}-6$ desaturase.

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