中文摘要：

运用SMART（Switching Mechanism at 5＇end of RNA Transcript）技术构建海南黄牛外周血白细胞cDNA文库。采用RNAiso Reagent处理新鲜血液，以总RNA抽提试剂盒（上海华舜）制备总RNA。用Powerscript^TM反转录酶逆转录合成第一链cDNA，LD-PCR扩增获得cDNA双链，SfiⅠ酶切和CHROMA SPIN-400^TM柱分离后，500bp以上的片段与pDNR—LIB载体连接，电转化入Ecoli．DH5α，建成原始文库。经鉴定，库容量达到1．2×10^6克隆，原始文库滴度为3×10^9cfu／mL，扩增后的文库滴度为4．3×10^10cfu／mL。PCR鉴定重组子，发现重组率接近100％，插入片段大小分布为0．5～2kb，插入片段平均长度约为1kb。文库的各项指标均达要求，为利用文库筛选海南黄牛已知和未知功能基因提供了材料来源，且为进一步研究基因的结构和功能奠定了重要基础。

英文摘要：

A cDNA Library from peripheral blood leucocytes in Hainan yellow cattle was constructed with the SMART （swithching mechanism at 5＇ end of RNA transcript） technique. Total RNA was extracted from periphead blood leucoytes and reversely transcripted into full-length cDNA by using Powerscript^TM. Double strand cDAN was synthesized and amplified by long-distance PCR （LD-PCR） . The PCR products were digested by proteinase K. After digestion with Sfi Ⅰ and size fraction using CHROMA SPIN-400^TM columns, cDNAs （〉500bp） were ligated to Sfi Ⅰ digested, dephosphorylated pDNR-LIB vector. The ligation mixture was transformed into Ecoli.DH5α by electroporation. The primarily constructed cDAN library contained 1.2×10^6 independent clones. The titer of cDNA library was estimated as 3×10^9 cfu/mL with the percentage of recombiant clones almost reaching 100%, and that of the amplified library was 6.3×10^10 cfu/mL. PCR results showed that the inserts varied from 0.5 to 2.0kb with average size being larger than 1000bp, which meets the requirement of a standard cDNA library. The construction of Hainan Yellow Cattle cDNA library could lay a basis for subsequent work of clone and selection of some kinds of fine functinal genes.