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211.
Yong Jia Caterina Selva Yujuan Zhang Bo Li Lee A. McFawn Sue Broughton Xiaoqi Zhang Sharon Westcott Penghao Wang Cong Tan Tefera Angessa Yanhao Xu Ryan Whitford Chengdao Li 《The Plant journal : for cell and molecular biology》2020,101(5):1057-1074
Functional divergence after gene duplication plays a central role in plant evolution. Among cereals, only Hordeum vulgare (barley), Triticum aestivum (wheat) and Secale cereale (rye) accumulate delphinidin‐derived (blue) anthocyanins in the aleurone layer of grains, whereas Oryza sativa (rice), Zea mays (maize) and Sorghum bicolor (sorghum) do not. The underlying genetic basis for this natural occurrence remains elusive. Here, we mapped the barley Blx1 locus involved in blue aleurone to an approximately 1.13 Mb genetic interval on chromosome 4HL, thus identifying a trigenic cluster named MbHF35 (containing HvMYB4H, HvMYC4H and HvF35H). Sequence and expression data supported the role of these genes in conferring blue‐coloured (blue aleurone) grains. Synteny analyses across monocot species showed that MbHF35 has only evolved within distinct Triticeae lineages, as a result of dispersed gene duplication. Phylogeny analyses revealed a shared evolution pattern for MbHF35 in Triticeae, suggesting that these genes have co‐evolved together. We also identified a Pooideae‐specific flavonoid 3′,5′‐hydroxylase (F3′5′H) lineage, termed here Mo_F35H2, which has a higher amino acid similarity with eudicot F3′5′Hs, demonstrating a scenario of convergent evolution. Indeed, selection tests identified 13 amino acid residues in Mo_F35H2 that underwent positive selection, possibly driven by protein thermostablility selection. Furthermore, through the interrogation of barley germplasm there is evidence that HvMYB4H and HvMYC4H have undergone human selection. Collectively, our study favours blue aleurone as a recently evolved trait resulting from environmental adaptation. Our findings provide an evolutionary explanation for the absence of blue anthocyanins in other cereals and highlight the importance of gene functional divergence for plant diversity and environmental adaptation. 相似文献
212.
Joe Carver Domingos Ng Michelle Zhou Peggy Ko Dejin Zhan Mandy Yim David Shaw Brad Snedecor Michael W. Laird Steven Lang Amy Shen Zhilan Hu 《Biotechnology progress》2020,36(4):e2967
Historically, therapeutic protein production in Chinese hamster ovary (CHO) cells has been accomplished by random integration (RI) of expression plasmids into the host cell genome. More recently, the development of targeted integration (TI) host cells has allowed for recombination of plasmid DNA into a predetermined genomic locus, eliminating one contributor to clone-to-clone variability. In this study, a TI host capable of simultaneously integrating two plasmids at the same genomic site was used to assess the effect of antibody heavy chain and light chain gene dosage on antibody productivity. Our results showed that increasing antibody gene copy number can increase specific productivity, but with diminishing returns as more antibody genes are added to the same TI locus. Random integration of additional antibody DNA copies in to a targeted integration cell line showed a further increase in specific productivity, suggesting that targeting additional genomic sites for gene integration may be beneficial. Additionally, the position of antibody genes in the two plasmids was observed to have a strong effect on antibody expression level. These findings shed light on vector design to maximize production of conventional antibodies or tune expression for proper assembly of complex or bispecific antibodies in a TI system. 相似文献
213.
Xiangming Li Yujian Zhang Li Jing Zongming Fu Ou Ma Jishna Ganguly Nilesh Vaidya Richard Sisson Jennifer Naginskaya Avinash Chinthala Minggang Cui Ryan Yamagata Mark Wilson Matthew Sanders Zihao Wang Paola Lo Surdo Marcin Bugno 《Biotechnology progress》2020,36(2):e2914
Mammalian cell line generation typically includes stable pool generation, single cell cloning and several rounds of clone selection based on cell growth, productivity and product quality criteria. Individual clone expansion and phenotype-based ranking is performed initially for hundreds or thousands of mini-scale cultures, representing the major operational challenge during cell line development. Automated cell culture and analytics systems have been developed to enable high complexity clone selection workflows; while ensuring traceability, safety, and quality of cell lines intended for biopharmaceutical applications. Here we show that comprehensive and quantitative assessment of cell growth, productivity, and product quality attributes are feasible at the 200–1,200 cell colony stage, within 14 days of the single cell cloning in static 96-well plate culture. The early cell line characterization performed prior to the clone expansion in suspension culture can be used for a single-step, direct selection of high quality clones. Such clones were comparable, both in terms of productivity and critical quality attributes (CQAs), to the top-ranked clones identified using an established iterative clone screening approach. Using a complex, multi-subunit antigen as a model protein, we observed stable CQA profiles independently of the cell culture format during the clonal expansion as well as in the batch and fed-batch processes. In conclusion, we propose an accelerated clone selection approach that can be readily incorporated into various cell line development workstreams, leading to significant reduction of the project timelines and resource requirements. 相似文献
214.
Ilse Van Gucht Josephina A.N. Meester Jotte Rodrigues Bento Maaike Bastiaansen Jarl Bastianen Ilse Luyckx Lotte Van Den Heuvel Cédric H.G. Neutel Pieter-Jan Guns Mandy Vermont Erik Fransen Melanie H.A.M. Perik Joe Davis Velchev Maaike Alaerts Dorien Schepers Silke Peeters Isabel Pintelon Abdulrahman Almesned Aline Verstraeten 《American journal of human genetics》2021,108(6):1115-1125
215.
Hui Li Shuyi Wang Fang-Hua Lee Ryan S. Roark Alex I. Murphy Jessica Smith Chengyan Zhao Juliette Rando Neha Chohan Yu Ding Eunlim Kim Emily Lindemuth Katharine J. Bar Ivona Pandrea Cristian Apetrei Brandon F. Keele Jeffrey D. Lifson Mark G. Lewis Thomas N. Denny Barton F. Haynes Beatrice H. Hahn George M. Shaw 《Journal of virology》2021,95(11)
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Pewowaruk Ryan Roldán-Alzate Alejandro 《Biomechanics and modeling in mechanobiology》2021,20(5):1659-1674
Biomechanics and Modeling in Mechanobiology - A distributed lumped parameter (DLP) model of blood flow was recently developed that can be simulated in minutes while still incorporating complex... 相似文献
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In Vitro Cellular & Developmental Biology - Plant - Micropropagation is a preferred method to propagate clean, clonal stock plants. Subculture is labor intensive and costly. In vitro hedging... 相似文献