【摘要】 通过序列设计、合成策略设计等方法 ,由CRL中最重要的同工酶LIP1的成熟多肽序列 ,生成适合在毕赤酵母中表达的基因序列sculipⅠ。通过链延伸反应及PCR反应将DNA单链人工合成为一段长为 16 35bp的DNA双链 ;并将其克隆入质粒pBluescriptⅡSK ( +)。PCR鉴定及测序结果表明sculipⅠ得到了正确的合成与克隆。还就若干长片段基因人工合成应该注意的问题进行讨论。更多还原

【Abstract】 According to the mature peptide sequence of LIP1, the most important isoenzyme of CRL, sculipⅠ, the gene sequence designed to be expressed in pichia pastria, was generated by methods of sequence and synthesis strategy design. 27 single-strain oligonucleotides were assembled as a 1 635 bp-length double-strains DNA after DNA strain extending and PCR reaction; the double-strains DNA was insert into plasmid pBluescript Ⅱ SK(+). PCR detection, sequence analysis showed the sculipⅠ was synthesized and cloned correctly. Some key points of synthesis of long double-strains DNA were also discussed.更多还原