TY - JOUR

T1 - The Structure of T-DNA Insertions in Transgenic Tobacco Plants Producing Bovine Interferon-Gamma

AU - Burlakovskiy, Mikhail

AU - Saveleva, Natalia

AU - Rumyantsev, Andrey M.

AU - Yemelyanov, Vladislav V.

AU - Padkina, Marina V.

AU - Lutova, Ludmila

N1 - Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2022/1/12

Y1 - 2022/1/12

N2 - Many of the most modern drugs are of a protein nature and are synthesized by transgenic producer organisms. Bacteria, yeast, or animal cell cultures are commonly used, but plants have a number of advantages—minimal biomass unit cost, animal safety (plants are not attacked by mammalian pathogens), the agricultural scale of production, and the ability to produce complex proteins. A disadvantage of plants may be an unstable level of transgene expression, which depends on the transgene structure and its insertion site. We analyzed the structure of T‐DNA inserts in transgenic tobacco plants (Nicotiana tabacum L.) belonging to two lines obtained using the same genetic construct but demonstrating different biological activities of the recombinant protein (bovine interferon‐gamma). We found that, in one case, T‐DNA was integrated into genomic DNA in the region of centromeric repeats, and in the other, into a transcriptionally active region of the genome. It was also found that in one case, the insert has a clustered structure and consists of three copies. Thus, the structure of T‐DNA inserts in both lines is not optimal (the optimal structure includes a single copy of the insert located in the active region of the genome). It is desirable to carry out such studies at the early stages of transgenic plants selection.

AB - Many of the most modern drugs are of a protein nature and are synthesized by transgenic producer organisms. Bacteria, yeast, or animal cell cultures are commonly used, but plants have a number of advantages—minimal biomass unit cost, animal safety (plants are not attacked by mammalian pathogens), the agricultural scale of production, and the ability to produce complex proteins. A disadvantage of plants may be an unstable level of transgene expression, which depends on the transgene structure and its insertion site. We analyzed the structure of T‐DNA inserts in transgenic tobacco plants (Nicotiana tabacum L.) belonging to two lines obtained using the same genetic construct but demonstrating different biological activities of the recombinant protein (bovine interferon‐gamma). We found that, in one case, T‐DNA was integrated into genomic DNA in the region of centromeric repeats, and in the other, into a transcriptionally active region of the genome. It was also found that in one case, the insert has a clustered structure and consists of three copies. Thus, the structure of T‐DNA inserts in both lines is not optimal (the optimal structure includes a single copy of the insert located in the active region of the genome). It is desirable to carry out such studies at the early stages of transgenic plants selection.

KW - Agrobacterial transformation

KW - Interferon‐gamma

KW - Nicotiana tabacum

KW - Transgenic plants

KW - T‐DNA insertion

KW - transgenic plants

KW - T-DNA insertion

KW - INTEGRATION

KW - SITES

KW - agrobacterial transformation

KW - PROTEINS

KW - interferon-gamma

UR - http://www.scopus.com/inward/record.url?scp=85122849776&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/db1dbce9-d21c-338a-9694-510b82d056d7/

U2 - 10.3390/app12020761

DO - 10.3390/app12020761

M3 - Article

VL - 12

JO - Applied Sciences (Switzerland)

JF - Applied Sciences (Switzerland)

SN - 2076-3417

IS - 2

M1 - 761

ER -