Novel GPS-containing G protein-coupled receptor from Monosiga brevicollis.
Novel GPS-containing G protein-coupled receptor from Monosiga brevicollis.
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来自 Monosiga brevicollis 的新型含 GPS 的 G 蛋白偶联受体。
DOI：
10.1134/s1607672909040061
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Petrenko,AG
中科院分区：
文献类型：
--
作者：
Serova,OV;Deyev,IE;Petrenko,AG
从M.通过聚合酶链反应（PCR）和5 '-cDNA末端快速扩增（5'-RACE）技术克隆了短颈霉（brevicollis）基因。根据来自M基因组数据库的预测基因序列estExt_fgenesh 1_pg. C_20766（蛋白质识别号35731）合成引物。短颈草v1. 0（http：//genome. jgipsf。org/Monbr 1/Monbr 1.回家html）。为了克隆方便，引物P1和P4含有在独特的NcoI限制性位点部分互补的序列;引物P3和P6含有在EcoRV限制性位点互补的序列。引物P2和P5分别与预测蛋白的C-和N-末端互补（表1，图1）。用引物P1/P2和P3/P4对单链cDNA进行PCR扩增，分别产生2100和1000 bp长的DNA片段。将这些片段克隆到如[7]所述从pUC 19载体（Fermentas）获得的T载体中，随后的测序证实了cDNA片段与基因组结构的对应性。然而，在用引物P5/P6进行PCR扩增后没有检测到反应产物。如[8]中所述，通过修饰的5 '-RACE确定蛋白质的N-末端序列。引物P7和P6分别用作第一和第二基因特异性引物。反应产生不同长度的DNA片段;将300-和360-bp产物克隆到T载体中并测序。这些程序允许
The full-length cDNA for the GPS-containing receptor from M. brevicollis was cloned by polymerase chain reaction (PCR) and rapid amplification of 5'-cDNA ends (5'-RACE). Primers were synthesized according to the predicted gene sequence estExt_fgenesh1_pg. C_20766 (protein identification number 35731) from the genome database for M. brevicollis v1. 0(http://genome. jgipsf. org/Monbr1/Monbr1. home. html). For cloning convenience, primers P1 and P4 contained the sequences that were partially complementary at the unique NcoI restriction site; primers P3 and P6, sequences complementary at the EcoRV restriction site. Primers P2 and P5 were complementary to the C-and N-termini, respectively, of the predicted protein (Table 1, Fig. 1).PCR amplification of single-stranded cDNA with primers P1/P2 and P3/P4 yielded DNA fragments 2100 and 1000 bp long, respectively. Cloning these fragments into the T-vector, which was obtained from the pUC19 vector (Fermentas) as described in [7], and subsequent sequencing confirmed the correspondence of cDNA fragments to the genome structure. However, no reaction products were detected after PCR amplification with the primers P5/P6. The N-terminal sequence of the protein was determined by the modified 5'-RACE as described in [8]. Primers P7 and P6 were used as the first and second gene-specific primers, respectively. The reaction yielded DNA fragments of different length; the 300-and 360-bp products were cloned into the T-vector and sequenced. These procedures allowed
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影响因子：
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作者：
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DOI：
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发表时间：
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期刊：
European journal of biochemistry
影响因子：
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作者：
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影响因子：
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作者：
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