Lou J, Manske P R, Aoki M, Joyce M E
Department of Orthopaedic Surgery, Washington University School of Medicine, St. Louis, Missouri, USA.
J Orthop Res. 1996 Jul;14(4):513-7. doi: 10.1002/jor.1100140403.
In this study, we successfully transferred the Escherichia coli beta-galactosidase gene. LacZ, into the chicken tendon and tendon sheath by a recombinant adenovirus. The recombinant adenovirus Adv-beta gal that carried the E. coli LacZ gene was constructed by homologous recombination in 293 cells (human transformed embryonic kidney) between the expressing vector and the ClaI large fragment of adenovirus 5 genome. Each chicken received a 10 microliters injection containing 10(5) plaque-forming units of recombinant virus Adv-beta gal. into the tendon sheath of the long toe Samples of tendon and tendon sheath were harvested at 3.30, and 75 days after the injection. The LacZ gene transfer was detected for its coding product beta-galactosidase by staining with X-gal solution. The results showed that all tendon and tendon sheath samples from the three harvest times stained positive (blue). The tendon sheath samples were more extensively stained; staining of the tendon was limited to the epitenon layer. These data suggest that a functional exogenous gene can potentially be transferred into the tendon and tendon sheath by similar techniques; such techniques may be used to improve healing and reduce adhesion formation.
在本研究中，我们通过重组腺病毒成功地将大肠杆菌β-半乳糖苷酶基因（LacZ）导入鸡的肌腱和腱鞘。携带大肠杆菌LacZ基因的重组腺病毒Adv-βgal是通过在293细胞（人转化胚胎肾细胞）中，表达载体与腺病毒5基因组的ClaI大片段之间的同源重组构建而成。每只鸡接受10微升含有10⁵个重组病毒Adv-βgal噬斑形成单位的注射，注射到长趾腱鞘中。在注射后3天、30天和75天采集肌腱和腱鞘样本。通过用X-gal溶液染色来检测LacZ基因转移产生的编码产物β-半乳糖苷酶。结果显示，在三个采集时间点采集的所有肌腱和腱鞘样本染色均呈阳性（蓝色）。腱鞘样本染色范围更广；肌腱的染色仅限于腱外膜层。这些数据表明，通过类似技术有可能将功能性外源基因导入肌腱和腱鞘；此类技术可用于促进愈合和减少粘连形成。
