المؤلفون: Liu, Hua Liang, Yin, Zhi Jie, Xiao, Li, Xu, Yi Nong, Qu, Le Qing

مصطلحات موضوعية: RESEARCH PAPER

الوصف: α-Linolenic acid (ALA) deficiency and a skewed of ω6:ω3 fatty acid ratio in the diet are a major explanation for the prevalence of cardiovascular diseases and inflammatory/autoimmune diseases. There is a need to enhance the ALA content and to reduce the ratio of linoleic acid (LA) to ALA. Six ω-3 (Δ-15) fatty acid desaturase (FAD) genes were cloned from rice and soybean. The subcellular localizations of the proteins were identified. The FAD genes were introduced into rice under the control of an endosperm-specific promoter, GluC , or a Ubi-1 promoter to evaluate their potential in increasing the ALA content in seeds. The ALA contents in the seeds of endoplasmic reticulum (ER)-localized GmFAD3-1 and OsFAD3 overexpression lines increased from 0.36 mg g−1 to 8.57 mg g−1and 10.06 mg g−1, respectively, which was 23.8- and 27.9-fold higher than that of non-transformants. The trait of high ALA content was stably inheritable over three generations. Homologous OsFAD3 is more active than GmFAD3-1 in catalysing LA conversion to ALA in rice seeds. Overexpression of ER-localized GmFAD3-2/3 and chloroplast-localized OsFAD7/8 had less effect on increasing the ALA content in rice seeds. The GluC promoter is advantageous compared with Ubi-1 in this experimental system. The enhanced ALA was preferentially located at the sn -2 position in triacylglycerols. A meal-size portion of high ALA rice would meet >80% of the daily adult ALA requirement. The ALA-rich rice could be expected to ameliorate much of the global dietary ALA deficiency.

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العلاقة: http://jxb.oxfordjournals.org/cgi/content/short/ers051v1Test; http://dx.doi.org/10.1093/jxb/ers051Test

الإتاحة: https://doi.org/10.1093/jxb/ers051Test
http://jxb.oxfordjournals.org/cgi/content/short/ers051v1Test

المؤلفون: Liu, Hua Liang, Yin, Zhi Jie, Xiao, Li, Xu, Yi Nong, Qu, Le Qing

مصطلحات موضوعية: Research Papers

الوصف: α-Linolenic acid (ALA) deficiency and a skewed of ω6:ω3 fatty acid ratio in the diet are a major explanation for the prevalence of cardiovascular diseases and inflammatory/autoimmune diseases. There is a need to enhance the ALA content and to reduce the ratio of linoleic acid (LA) to ALA. Six ω-3 (Δ-15) fatty acid desaturase (FAD) genes were cloned from rice and soybean. The subcellular localizations of the proteins were identified. The FAD genes were introduced into rice under the control of an endosperm-specific promoter, GluC , or a Ubi-1 promoter to evaluate their potential in increasing the ALA content in seeds. The ALA contents in the seeds of endoplasmic reticulum (ER)-localized GmFAD3-1 and OsFAD3 overexpression lines increased from 0.36 mg g−1 to 8.57 mg g−1and 10.06 mg g−1, respectively, which was 23.8- and 27.9-fold higher than that of non-transformants. The trait of high ALA content was stably inheritable over three generations. Homologous OsFAD3 is more active than GmFAD3-1 in catalysing LA conversion to ALA in rice seeds. Overexpression of ER-localized GmFAD3-2/3 and chloroplast-localized OsFAD7/8 had less effect on increasing the ALA content in rice seeds. The GluC promoter is advantageous compared with Ubi-1 in this experimental system. The enhanced ALA was preferentially located at the sn -2 position in triacylglycerols. A meal-size portion of high ALA rice would meet >80% of the daily adult ALA requirement. The ALA-rich rice could be expected to ameliorate much of the global dietary ALA deficiency.

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العلاقة: http://jxb.oxfordjournals.org/cgi/content/short/63/8/3279Test; http://dx.doi.org/10.1093/jxb/ers051Test

الإتاحة: https://doi.org/10.1093/jxb/ers051Test
http://jxb.oxfordjournals.org/cgi/content/short/63/8/3279Test

المؤلفون: Yin, Chang-Cheng, Ren, Li-Li, Zhu, Lin-Lin, Wang, Xiang-Bin, Zhang, Zhong, Huang, Hua-Liang, Yan, Xi-Yun

مصطلحات موضوعية: Regular Paper

الوصف: The randomsation scheme of hypervariable region takes crucial roles in construction of a synthetic antibody library. The codon bias and inevitable ‘stop’ codon of conventional “NNK” and “NNS” codons limit their applications. Here we report a split-mix-split DNA synthesis method that can control over the amino acid composition and distribution of randomized sequences effectually. A fully synthetic human antibody library with a diversity of 1.56×109 was successfully generated with complementarity determining region 3 (CDR3) randomized by this strategy. Sequencing analysis indicated that more than 60% of colonies had completely correct scFv genes and the amino acid composition and distribution were accordance with design well. The utility was demonstrated by screening of scFv clones against BHL (anti-CD3×anti-carcinoma bispecific antibody). These results proved the feasibility of the split-mix-split DNA randomsation strategy in library construction and site-directed mutagenesis.

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العلاقة: http://jb.oxfordjournals.org/cgi/content/short/mvn103v1Test; http://dx.doi.org/10.1093/jb/mvn103Test

الإتاحة: https://doi.org/10.1093/jb/mvn103Test
http://jb.oxfordjournals.org/cgi/content/short/mvn103v1Test

المؤلفون: Yin, Chang-Cheng, Ren, Li-Li, Zhu, Lin-Lin, Wang, Xiang-Bin, Zhang, Zhong, Huang, Hua-Liang, Yan, Xi-Yun

مصطلحات موضوعية: Regular Papers

الوصف: The randomization scheme of hypervariable region takes crucial role in construction of a synthetic antibody library. The codon bias and inevitable ‘stop’ codon of conventional ‘NNK’ and ‘NNS’ codons limit their applications. Here we report a split–mix–split DNA synthesis method that can control over the amino acid composition and distribution of randomized sequences effectually. A fully synthetic human antibody library with a diversity of 1.56 × 109 was successfully generated with complementarity determining region 3 (CDR3) randomized by this strategy. Sequencing analysis indicated that >60% of colonies had completely correct scFv genes and the amino acid composition and distribution were designed well in accordance. The utility was demonstrated by screening of scFv clones against BHL (anti-CD3 × anti-ovarian carcinoma bispecific antibody). These results proved the feasibility of the split–mix–split DNA randomization strategy in library construction and site-directed mutagenesis.

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العلاقة: http://jb.oxfordjournals.org/cgi/content/short/144/5/591Test; http://dx.doi.org/10.1093/jb/mvn103Test

الإتاحة: https://doi.org/10.1093/jb/mvn103Test
http://jb.oxfordjournals.org/cgi/content/short/144/5/591Test

المؤلفون: HUANG, ZHAN-PENG, ZHOU, HUI, HE, HUA-LIANG, CHEN, CHUN-LONG, LIANG, DAN, QU, LIANG-HU

مصطلحات موضوعية: ARTICLE

الوصف: Small nucleolar RNAs (snoRNAs) are an abundant group of noncoding RNAs mainly involved in the post-transcriptional modifications of rRNAs in eukaryotes. In this study, a large-scale genome-wide analysis of the two major families of snoRNA genes in the fruit fly Drosophila melanogaster has been performed using experimental and computational RNomics methods. Two hundred and twelve gene variants, encoding 56 box H/ACA and 63 box C/D snoRNAs, were identified, of which 57 novel snoRNAs have been reported for the first time. These snoRNAs were predicted to guide a total of 147 methylations and pseudouridylations on rRNAs and snRNAs, showing a more comprehensive pattern of rRNA modification in the fruit fly. With the exception of nine, all the snoRNAs identified to date in D. melanogaster are intron encoded. Remarkably, the genomic organization of the snoRNAs is characteristic of 8 dUhg genes and 17 intronic gene clusters, demonstrating that distinct organizations dominate the expression of the two families of snoRNAs in the fruit fly. Of the 267 introns in the host genes, more than half have been identified as host introns for coding of snoRNAs. In contrast to mammals, the variation in size of the host introns is mainly due to differences in the number of snoRNAs they contain. These results demonstrate the extensive utilization of introns for coding of snoRNAs in the host genes and shed light on further research of other noncoding RNA genes in the large introns of the Drosophila genome.

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العلاقة: http://rnajournal.cshlp.org/cgi/content/short/11/8/1303Test; http://dx.doi.org/10.1261/rna.2380905Test

الإتاحة: https://doi.org/10.1261/rna.2380905Test
http://rnajournal.cshlp.org/cgi/content/short/11/8/1303Test

المؤلفون: HUANG, ZHAN-PENG, ZHOU, HUI, HE, HUA-LIANG, CHEN, CHUN-LONG, LIANG, DAN, QU, LIANG-HU

مصطلحات موضوعية: ARTICLE

الوصف: Small nucleolar RNAs (snoRNAs) are an abundant group of noncoding RNAs mainly involved in the post-transcriptional modifications of rRNAs in eukaryotes. In this study, a large-scale genome-wide analysis of the two major families of snoRNA genes in the fruit fly Drosophila melanogaster has been performed using experimental and computational RNomics methods. Two hundred and twelve gene variants, encoding 56 box H/ACA and 63 box C/D snoRNAs, were identified, of which 57 novel snoRNAs have been reported for the first time. These snoRNAs were predicted to guide a total of 147 methylations and pseudouridylations on rRNAs and snRNAs, showing a more comprehensive pattern of rRNA modification in the fruit fly. With the exception of nine, all the snoRNAs identified to date in D. melanogaster are intron encoded. Remarkably, the genomic organization of the snoRNAs is characteristic of 8 dUhg genes and 17 intronic gene clusters, demonstrating that distinct organizations dominate the expression of the two families of snoRNAs in the fruit fly. Of the 267 introns in the host genes, more than half have been identified as host introns for coding of snoRNAs. In contrast to mammals, the variation in size of the host introns is mainly due to differences in the number of snoRNAs they contain. These results demonstrate the extensive utilization of introns for coding of snoRNAs in the host genes and shed light on further research of other noncoding RNA genes in the large introns of the Drosophila genome.

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العلاقة: http://www.rnajournal.org/cgi/content/short/rna.2380905v1Test; http://dx.doi.org/10.1261/rna.2380905Test

الإتاحة: https://doi.org/10.1261/rna.2380905Test
http://www.rnajournal.org/cgi/content/short/rna.2380905v1Test

المؤلفون: Wang, Xiang-Bin, Zhao, Bao-Feng, Zhao, Qi, Piao, Jin-Hua, Liu, Jing, Lin, Qing, Huang, Hua-Liang

مصطلحات موضوعية: BIOTECHNOLOGY

الوصف: Anti-tumor associated antigen (TAA)⋅CD3⋅CD28 trispecific antibody(TsAb) is able to provide two signals for fully and continuously activation of T lymphocytes and recruit them around tumor cells, presenting an attractive concept in tumor immunotherapy. Here, a new single chain trispecific antibody (scTsAb), named CEA-scTsAb, was constructed by fusion of anti-CEA (Carcinoma Embryonic Antigen) single chain antibody (scFv), anti-CD3 scFv and anti-CD28 VH, spaced by polypeptide interlinkers taken from the fragment of constant region (FC) of human IgG and human serum albumin (HSA). It was expressed in Escherichia coli at low temperature (30°C) with up to 50% of the antibody being present in soluble form. After one step of DEAE anion chromatography, the soluble product was sufficiently pure for further in vitro activity assays. First, it was proved that CEA-scTsAb could recognize three antigens (CEA, CD28 and Jurkat cell membrane antigen) specifically and could distinguish antigen positive cells from antigen negative cells in vitro . Then fresh PBMC (peripheral blood mononuclear cells), without being pre-treated by co-stimulatory reagents, such as IL-2 or CD28 mAb, were used as effector cells to test their ability to mediate tumor specific cytolysis of CEA-positive tumor cells, SW1116. It was found by photomicrography that T lymphocytes were attracted to SW1116 cells in the presence of CEA-scTsAb, which resulted in effective cytolysis of tumor cells. As shown by MTT assay, the efficacy of tumor specific cytolysis mediated by CEA-scTsAb related to both the quantity and activation of PBMC. At an effector cells/target cells ratio (E/T) of 5, it was proved by dual-color FACS with propidium iodide (PI) and FITC-annexin V that both necrosis and apoptosis of tumor cells were causes of tumor specific cytolysis. In summary, a new single chain trispecific (CEA × CD3 × CD28) antibody was constructed and characterized carefully in this paper and was found to possess functions: (i) to activate T lymphocytes independently of ...

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العلاقة: http://jb.oxfordjournals.org/cgi/content/short/135/4/555Test; http://dx.doi.org/10.1093/jb/mvh065Test

الإتاحة: https://doi.org/10.1093/jb/mvh065Test
http://jb.oxfordjournals.org/cgi/content/short/135/4/555Test

المؤلفون: Chun, Lei, Yin, Chang-Cheng, Song, Jing-Zhen, Liu, Ming-Xue, Piao, Jin-Hua, Lin, Qing, Wang, Xiang-Bin, Huang, Hua-Liang

مصطلحات موضوعية: MOLECULAR BIOLOGY

الوصف: Secondary lymphoid tissue chemokine (SLC) is a CC chemokine that plays an important role in leukocytes homing to lymphoid tissues. The ability of SLC to co-localize both T cells and dendritic cells formed the rationale to evaluate its utility in cancer immunotherapy. The in vivo antitumor effect of murine SLC (mSLC) has been well documented, but little is known about that of human SLC (hSLC). To investigate the antitumor efficiency in vivo of hSLC, the hSLC gene was artificially synthesized and induced to express as a soluble form in Escherichia coli . After purification, the purity of the recombinant human SLC (rhSLC) protein was above 95% by SDS-PAGE analysis. The K d of rhSLC binding to peripheral blood lymphocytes (PBLs) was 0.2186 ± 0.02675 µM as assessed by FACS, and the maximal chemotactic index of rhSLC was 9.49 at 100 nM as assessed by in vitro chemotaxis assay. Then genomic sequences of hSLC and mSLC, and of human CCR7 (hCCR7) and murine CCR7 (mCCR7), the receptor for SLC, were aligned. It was found that hSLC and mSLC share 70.72% identity and hCCR7 and mCCR7share 86.77% identity. Furthermore, we found that rhSLC could chemoattract murine peripheral blood mononuclear cells (PBMCs) in vitro . On the basis of these facts, immune competent mice inoculated with S180 sarcoma cells were chosen as an in vivo model. Intratumoral injections of rhSLC inhibited tumor growth and increased survival. These findings suggest that, despite its incapability to bind to either human or murine CXCR3, which is related to angiostasis, rhSLC can induce an antitumor response in vivo by another route. This report proves that rhSLC has a potent tumor-inhibition ability that makes it a promising candidate agent in cancer immunotherapy.

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العلاقة: http://jb.oxfordjournals.org/cgi/content/short/136/6/769Test; http://dx.doi.org/10.1093/jb/mvh186Test

الإتاحة: https://doi.org/10.1093/jb/mvh186Test
http://jb.oxfordjournals.org/cgi/content/short/136/6/769Test

المؤلفون: Wang, Xiang-Bin, Zhou, Bing, Yin, Chang-Cheng, Lin, Qing, Huang, Hua-Liang

مصطلحات موضوعية: BIOTECHNOLOGY

الوصف: Antibody reshaping is an effective way to reduce the immunogenicity while maintaining or improving the affinity of murine antibodies. This paper describe a new in vitro approach for rapidly reshaping murine antibodies by combining DNA shuffling with ribosome display. With the new method, a reshaping anti–4-1BB single-chain antibody (scFv), Re-4B4-1 scFv, which bound to its antigen (4-1BB) specifically and strongly, was selected from a reshaping library. These results proved definitely the feasibility of the new designed approach for antibody reshaping.

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العلاقة: http://jb.oxfordjournals.org/cgi/content/short/136/1/19Test; http://dx.doi.org/10.1093/jb/mvh083Test

الإتاحة: https://doi.org/10.1093/jb/mvh083Test
http://jb.oxfordjournals.org/cgi/content/short/136/1/19Test