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},{"id":"R640","pred":"conj","subj":"T1138","obj":"T1134"},{"id":"R641","pred":"compound","subj":"T1139","obj":"T1138"},{"id":"R642","pred":"punct","subj":"T1140","obj":"T1125"},{"id":"R643","pred":"advmod","subj":"T1141","obj":"T1125"},{"id":"R644","pred":"punct","subj":"T1142","obj":"T1104"},{"id":"R391","pred":"pobj","subj":"T867","obj":"T865"},{"id":"R397","pred":"dobj","subj":"T875","obj":"T872"},{"id":"R402","pred":"nsubjpass","subj":"T880","obj":"T873"},{"id":"R413","pred":"dobj","subj":"T891","obj":"T889"},{"id":"R498","pred":"pobj","subj":"T984","obj":"T982"},{"id":"R509","pred":"nmod","subj":"T997","obj":"T1000"},{"id":"R521","pred":"dobj","subj":"T1009","obj":"T995"},{"id":"R528","pred":"pobj","subj":"T1016","obj":"T1014"},{"id":"R545","pred":"dobj","subj":"T1035","obj":"T1033"}],"text":"Molecular cloning of the Acdp gene family\nTo clone the mouse Acdp genes, the human ACDP cDNA and predicted protein sequences were used to search the mouse EST database with the blastn and tblastn programs. Mouse EST markers corresponding to each Acdp member were then identified. For example, EST H3086H12-5 corresponds to Acdp1, W98010 for Acdp2, 603299135F1 for Acdp3 and BG083791 for Acdp4. A modified oligo-dT with a M13 tail was used for the RT reaction. A forward primer from each EST marker and the M13 primer (olig-dT tail) were used to amplify the 3' UTR sequence for each corresponding Acdp gene from the RT products. To obtain 5'-end coding sequences for the Acdp genes, we conducted a series nested PCR with combinations of mouse and human primers. The 5' UTR sequences were identified by directly sequencing BAC DNA containing the corresponding Acdp genes. The BAC clones were identified by screening a CITB mouse BAC DNA library (Research Genetics). The 5' UTR sequences obtained from above were further confirmed by RT-PCR. The Acdp1 gene contains 3,631 bp of nucleotide sequence and encodes a predicted protein with 951 amino acids (AA). The other three Acdp genes (Acdp2, 3 and 4) contain 3,244 bp, 2,684 bp and 2,743 bp of cDNA sequences, and encode deduced proteins of 874 amino acids, 713 amino acids and 771 amino acids, respectively."}
craft-ca-core-dev
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craft-ca-core-ex-dev
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cloning of the Acdp gene family\nTo clone the mouse Acdp genes, the human ACDP cDNA and predicted protein sequences were used to search the mouse EST database with the blastn and tblastn programs. Mouse EST markers corresponding to each Acdp member were then identified. For example, EST H3086H12-5 corresponds to Acdp1, W98010 for Acdp2, 603299135F1 for Acdp3 and BG083791 for Acdp4. A modified oligo-dT with a M13 tail was used for the RT reaction. A forward primer from each EST marker and the M13 primer (olig-dT tail) were used to amplify the 3' UTR sequence for each corresponding Acdp gene from the RT products. To obtain 5'-end coding sequences for the Acdp genes, we conducted a series nested PCR with combinations of mouse and human primers. The 5' UTR sequences were identified by directly sequencing BAC DNA containing the corresponding Acdp genes. The BAC clones were identified by screening a CITB mouse BAC DNA library (Research Genetics). The 5' UTR sequences obtained from above were further confirmed by RT-PCR. The Acdp1 gene contains 3,631 bp of nucleotide sequence and encodes a predicted protein with 951 amino acids (AA). The other three Acdp genes (Acdp2, 3 and 4) contain 3,244 bp, 2,684 bp and 2,743 bp of cDNA sequences, and encode deduced proteins of 874 amino acids, 713 amino acids and 771 amino acids, respectively."}