Poly (A) RNA was isolated from microdissected guinea pig organ of Corti and converted into cDNA with RNase H- murine leukemia virus reverse transcriptase. After size fractionation, the cDNA was directionally ligated into the vector pSPORT 1 and the plasmids were transformed into DH10B E. coli via electroporation. The library was found to have 3.35 x 10(6) independent colonies with ten percent of the colonies lacking an insert. After checking 33 randomly selected colonies for inserts, the average insert size was 1218 base pairs, ranging from 3300 base pairs to 400 base pairs. The library was screened with a beta-actin oligonucleotide probe and 1.4% of the colonies contained an insert hybridizing to the probe.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1016/0378-5955(92)90208-5 | DOI Listing |
Nat Commun
January 2025
Department of Physics and Astronomy, Michigan State University, East Lansing, MI, USA.
DEAD-box RNA-dependent ATPases are ubiquitous in all domains of life where they bind and remodel RNA and RNA-protein complexes. DEAD-box ATPases with helicase activity unwind RNA duplexes by local opening of helical regions without directional movement through the duplexes and some of these enzymes, including Ded1p from Saccharomyces cerevisiae, oligomerize to effectively unwind RNA duplexes. Whether and how DEAD-box helicases coordinate oligomerization and unwinding is not known and it is unclear how many base pairs are actively opened.
View Article and Find Full Text PDFJ Phys Chem B
January 2025
Department of Physics of Complex Systems, S. N. Bose National Centre for Basic Sciences, Block-JD, Sector-III, Salt Lake, Kolkata 700106, India.
In DNA double helices, Hoogsteen (HG) base pairing is an alternative mode of Watson-Crick (WC) base pairing. HG bp has a different hydrogen bonding pattern than WC bp. We investigate here the binding energy of homeodomain proteins with a HG-DNA duplex, where DNA adopts a HG bp in its sequence.
View Article and Find Full Text PDFPathogens
January 2025
Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Center for Disease Control and Prevention, Jiaxing 314050, China.
Mpox, a zoonotic disease caused by the mpox virus (MPXV), has seen a significant shift in its epidemiological status since 2022, evolving from an initial local outbreak to a global epidemic. This recent outbreak of MPXV mainly emerged in several European and American countries and subsequently spread to over 100 countries and regions worldwide. The rapid evolution of MPXV, coupled with increased international interactions, has led to a gradual rise in mpox cases in certain regions of Asia, mostly involving MPXV clade II and its branch strains.
View Article and Find Full Text PDFInt J Mol Sci
January 2025
Biotechnology Department, Sirius University of Science and Technology, 354349 Sirius, Russia.
In addition to the 20 canonical amino acids encoded in the genetic code, there are two non-canonical ones: selenocysteine and pyrrolysine. The discovery of pyrrolysine synthetases (PylRSs) was a key event in the field of genetic code expansion research. The importance of this discovery is mainly due to the fact that the translation systems involving PylRS, pyrrolysine tRNA (tRNA) and pyrrolysine are orthogonal to the endogenous translation systems of organisms that do not use this amino acid in protein synthesis.
View Article and Find Full Text PDFMicromachines (Basel)
January 2025
State Key Laboratory of Radio Frequency Heterogeneous Integration, Shanghai Jiao Tong University, Shanghai 200240, China.
On-chip gene synthesis has the potential to improve the synthesis throughput and reduce the cost exponentially. While there exist several microarray-based oligo synthesis technologies, on-chip gene assembly has yet to be demonstrated. This work introduces a novel on-chip DNA assembly method via dielectrophoresis (DEP) that can potentially be integrated with microarray-based oligo synthesis on the same chip.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!