RT-PCR and RACE techniques were used to clone 3＇ end real sequence of RPL11 gene from a goat. The sequences included some non-structural protein （3D） genes, and 3D gene immediately downstream 3＇ non-coding region ...RT-PCR and RACE techniques were used to clone 3＇ end real sequence of RPL11 gene from a goat. The sequences included some non-structural protein （3D） genes, and 3D gene immediately downstream 3＇ non-coding region （UTR） and poly （A） tail. The results showed that the length of specific amplified fragment was 566 bp （not including polyA tail）. RPL11 gene encoded 188 amino acids, amino acid sequence was conducted for homology analysis by NCBI BLAST tool. The highest homology was Mus （RPL11, mRNA homology 99%）, followed by Bos （RPL11, mRNA homology 96%）, and the next was Hs （RPL11, mRNA homology of 93%）.展开更多
Total RNA was isolated from mycelium of T. harzianum by Total RNA extraction kit, and two clear bands of rRNA （28S and 18S） were observed in agarose electrophoresis. By joining the 3＇end sequence with the known SA7...Total RNA was isolated from mycelium of T. harzianum by Total RNA extraction kit, and two clear bands of rRNA （28S and 18S） were observed in agarose electrophoresis. By joining the 3＇end sequence with the known SA76 EST from cDNA library of T. harzianum, a full-length cDNA sequence of 2019bp was obtained, whose open reading frame contained 1593bp, a stop codon TAA, a 5＇untranslated region （5＇UTR） of 266bp, a 3＇untranslated region （3＇UTR） of 201bp, and poly （A） 29 encoded a protein of 530 amino acids, had a signal peptide. T. harzianum shared 53% identity of secreted aspartic proteinase gene with G. zeae, 37% with N. crassa and 36% with C. globosum. The full-length cDNA sequence of secreted aspartic proteinase gene from T. harzianum was cloned for the first time by using BD SMART RACE technique, which provides a foundation to obtain and validate functional genes of T. harzianum.展开更多
基金Supported by National High-tech Research and Development Program of China (863 Program) (2006AA10Z1A4)International Cooperation Projects of Heilongjiang Province (WB07A06)Innovation Team Project of Northeast Agricultural University (CXT005-1-1)
文摘RT-PCR and RACE techniques were used to clone 3＇ end real sequence of RPL11 gene from a goat. The sequences included some non-structural protein （3D） genes, and 3D gene immediately downstream 3＇ non-coding region （UTR） and poly （A） tail. The results showed that the length of specific amplified fragment was 566 bp （not including polyA tail）. RPL11 gene encoded 188 amino acids, amino acid sequence was conducted for homology analysis by NCBI BLAST tool. The highest homology was Mus （RPL11, mRNA homology 99%）, followed by Bos （RPL11, mRNA homology 96%）, and the next was Hs （RPL11, mRNA homology of 93%）.
基金This paper was supported by Development Program (863) of China (No. 2003AA241140)
文摘Total RNA was isolated from mycelium of T. harzianum by Total RNA extraction kit, and two clear bands of rRNA （28S and 18S） were observed in agarose electrophoresis. By joining the 3＇end sequence with the known SA76 EST from cDNA library of T. harzianum, a full-length cDNA sequence of 2019bp was obtained, whose open reading frame contained 1593bp, a stop codon TAA, a 5＇untranslated region （5＇UTR） of 266bp, a 3＇untranslated region （3＇UTR） of 201bp, and poly （A） 29 encoded a protein of 530 amino acids, had a signal peptide. T. harzianum shared 53% identity of secreted aspartic proteinase gene with G. zeae, 37% with N. crassa and 36% with C. globosum. The full-length cDNA sequence of secreted aspartic proteinase gene from T. harzianum was cloned for the first time by using BD SMART RACE technique, which provides a foundation to obtain and validate functional genes of T. harzianum.
