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Studies on cell suspension Culture of Ricinus communis L.

Agricultural Biotechnology 2013年第6期2卷 21-23,28页

作者： Wei KANG Jin ZHENG Hubei Polytechnic University Hubei Key Laboratory of Mine Environmental Pollution Control & Remediation

Ricinus communis L. is a new copper hyperaccumulator growing on Tonglushan copper mine in Hubei Province, China. This study aimed to establish a suspension cell line of R. communis L. with stable and rapid growth for further screening of heavy metal-resistant R. communis L. cells and breeding of hyperaccu- mulators. In this study, cell suspension culture conditions were optimized by using orthogonal experimental design with previously induced R. communis L. embyre- genic calluses as experimental materials, to establish the suspension cell line of R. communis L. Under the optimal conditions, growth curves of suspension cells and changes in pH of culture liquid were determined. The results showed that the optimal culture conditions for R. conmmnis L. suspension cell line were ： MS ＋ O. 5 rag/I, 6-BA ＋ O. 2 mg/L NAA ＋ 50 mg/L sucrose ＋ 350 mg/L casein hydrolysate as basic medium, with callus inoculation amount of 2.5 g/50 ml, dark culture at （26 ±2） ℃ with shaking at 110 r/min. Under these conditions, increment of fresh weight and dry weight ofR. commun/s L. suspension cells reached the maximum of 4.56 g/（50 ml 14 d） and 0.49 g/（50 ml 16 d）, respectively. Growth curves of R. communis L. suspension cells were basically in ＂S＂ shape. Each culture cycle lasted 16 d, and the rapid growth stage was from the 6th d to the 14th d. In a culture cycle, pH of the culture liquid declined first and then increased to the maximum and stabilized gradually.

关键词： Ricinus communis L. Embyrogenic callus Orthogonal experimental design suspension cell line

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A Deleted Deletion Site in a New Vector Strain and Exceptional Genomic Stability of Plaque-Purified Modified Vaccinia Ankara(MVA)

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Virologica Sinica 2020年第2期35卷 212-226页

作者： Ingo Jordan Deborah Horn Kristin Thiele Lars Haag Katharina Fiddeke Volker Sandig ProBioGen AG.Herbert-Bayer-StraBe 8 13086 BerlinGermany Sartorius Stedim cellca GmbH.Erwin-Rentschler-Str 21 88471 LaupheimGermany Vironova AB Giivlegatan 2211330 StockholmSweden Department of Laboratory Medicine Karolinska Universitetsjukhuset i Huddinge14152 HuddingeSweden

Vectored vaccines based on highly attenuated modified vaccinia Ankara(MVA) are reported to be immunogenic, tolerant to pre-existing immunity, and able to accommodate and stably maintain very large transgenes. MVA is usually produced on primary chicken embryo fibroblasts, but production processes based on continuous cell lines emerge as increasingly robust and cost-effective alternatives. An isolate of a hitherto undescribed genotype was recovered by passage of a nonplaque-purified preparation of MVA in a continuous anatine suspension cell line(***) in chemically defined *** novel isolate(MVA-CR19) replicated to higher infectious titers in the extracellular volume of suspension cultures and induced fewer syncytia in adherent cultures. We now extend previous studies with the investigation of the point mutations in structural genes of MVA-CR19 and describe an additional point mutation in a regulatory gene. We furthermore map and discuss an extensive rearrangement of the left telomer of MVA-CR19 that appears to have occurred by duplication of the right telomer. This event caused deletions and duplications of genes that may modulate immunologic properties of MVACR19 as a vaccine vector. Our characterizations also highlight the exceptional genetic stability of plaque-purified MVA:although the phenotype of MVA-CR19 appears to be advantageous for replication, we found that all genetic markers that differentiate wildtype and MVA-CR19 are stably maintained in passages of recombinant viruses based on either wildtype or MVA-CR.

关键词： Modified vaccinia Ankara(MVA) MVA-CR19 CR.pIX suspension cell line Vaccine production Genome stability

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