Year-end Sale % OFF 
Abstract
Recombinant clones containing cDNA sequences coding for human apolipoprotein CII have been isolated from a human adult liver cDNA library. The clones were detected by hybridization with a mixed probe consisting of 96 different oligonucleotides 17 bases long, complementary to the mRNA coding for amino acids 66 to 71 of apolipoprotein CII. The largest cDNA insert is 439 nucleotides long and contains the complete sequence for the mature protein, the 3' untranslated sequence, and sequence coding for 17 amino acids of a signal peptide. The cDNA clone was used to detect apolipoprotein CII mRNA approximately 500 bases long in human liver and intestine RNA.
Highlights
After completion of the synthesis the oligonucleotides were purified by anion exchange and reverse phase high pressure liquid chromatography [17]
The mixed probe constructed by us (Fig. 2)contained oligonucleotides complementary to all possible combinations of codons for amino acids 66 to 71 of apo-CII.?Because one of the 96 different sequences would match the mRNA sequence exactly, high stringency washescould be used toeliminatethedetection of false positives during the screening[12, 27]
Preliminaryrestrictionmapping showed that allclones contained a cDNA insert which could be excised with PstI
Summary
Recombinantclones containing cDNA sequences cod-clones were plated at a density of 500 colonies per 82-mm diametering for human apolipoprotein CII have been isolated fromahumanadult liver cDNA library. At each position of ambiguity predicted by the amino signal peptide.ThecDNAclone was usedtodetect acid sequence equimolar mixtures of the appropriate dimers were apolipoprotein CIImRNA approximately 500 bases long in human liver and intestine RNA. Deficient patientsatthe molecular level, we have isolated cDNA recombinants containing coding sequences for apo-CII. The 5’end of the cDNA wascloned in reverse orientation by isolation of the Screening of cDNAClones-The construction of the adult liver HinfI fragment A of clone G24.After filling in of the ends with cDNA library has been described previously [12]. The relative intensities of the bands detected were determined by scanningtheautoradiogramswith a densitometer(Helena Quick Scan)
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
