A monoclonal antibody (2F4) directed against a 32-kDa dense-granule antigen of Sarcocystis muris cyst merozoites (bradyzoites) was used to screen a lambda ZAP cDNA expression library. A clone with an insert of 1.4 kb in length (DG 32/1) was isolated. A fusion protein derived from bacteria harbouring the recombinant plasmid DG 32/1 reacted with monoclonal antibody (mAb) 2F4. Southern blot hybridization suggests that the gene is present as a single copy. On Northern blots, a single mRNA species of 1.8 kb was detected by a cDNA-derived probe. In addition, we isolated a full-length clone (DG 32/PH1) by screening the cDNA library with a non-radioactive-labelled cDNA probe. The nucleotide sequence of DG 32/PH1 comprises 1.57 kb. It contains an open reading frame of 882 bp with a coding capacity of approximately 32 kDa. The hypothetical polypeptide consists of a putative N-terminal signal peptide and the mature protein sequence. The occurrence of an N-terminal signal sequence is consistent with the observation that the 32-kDa protein of S. muris is secreted from the dense granules.
Here, we present the nucleotide sequence of the bacteriophage T5 EcoRl/HpaI DNA fragment (27.8%-29.5% Rhoades' map (1)). The fragment contains six tRNA genes. Earlier corresponding transfer RNAs were identified by Hunt et al. (2), and tRNAAs was sequenced (3). Except tRNA genes four ORFs were found, and three of them have suitable RBS which are located on the proper distance from the initiation codons. The promoter sequences upstream the genes for tRNATYr and ORF 42aa were characterized by the cloning in the specialized promoter-probe plasmid pKWIII (the data will be published elsewhere). EMBL accession no. Z14121 Noncoding (RNA-like) strand is presented in the direction 5'-3' from the HpaI site. This sequence is immediately adjacent to the distal part of T5 tRNA gene region we sequenced earlier (4).
2,2,4,4‐Tetramethyl‐l ,3‐cyclopentandion (I) wird zum Alkohol (II) reduziert und zu (IIIa) dehydratisiert, aus dem das Hydrazon (IIIb), die Diazoverbindung (IIIc) sowie das Thion (IIId) hergestellt werden können.
Reduction of the chlorophenylisoquinolinecarbaldehyde (I) with sodium borohydride yields the alcohol (II) which is dehalogenated to produce the hydroxymethylphenylisoquinoline (III).
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